Chapter 20 Biotechnology
|
|
- Gerald Joel Powers
- 5 years ago
- Views:
Transcription
1 Chapter 20 Biotechnology
2 Manipulation of DNA In 2007, the first entire human genome had been sequenced. The ability to sequence an organisms genomes were made possible by advances in biotechnology, (the manipulation of organisms or their components to make useful products) and genetic engineering (the direct manipulation of genes for practical purposes). Recombinant DNA, DNA molecules formed when segments of DNA from two different sources (often different species) are combined in vitro.
3 Concept 20.1 DNA cloning yields multiple copies of a gene or other DNA segments DNA cloning can be used to amplify DNA sequences of interest Restriction enzymes can be used to insert foreign DNA into bacterial plasmids which can then be transformed into bacteria to produce recombinant bacteria
4 DNA Cloning Recall: The E. coli chromosome is a large circular molecule of DNA. In addition, bacteria may also contain plasmids, small circular DNA molecules. In gene cloning, multiple copies of a single gene are produced. To clone DNA, plasmids can be isolated from a bacterial cell and then foreign DNA can be inserted to create a recombinant DNA molecule. This is inserted back into the bacterium to generate a recombinant bacterium which will undergo cell division to produce more recombinant bacteria.
5 Applications of DNA Cloning A bacterial plasmid is a cloning vector a DNA molecule that can carry foreign DNA into a host cell and replicate there. Foreign DNA can be inserted into the cloning vector to generate a recombinant plasmid Two purposes of gene cloning: Make many copies of a gene Produce protein product Both of these can be isolated and harvested for use in research. They also have applications purposes in many industries E.g. agriculture, medicine
6 Bacterium 1 Gene inserted into plasmid Cell containing gene of interest Bacterial chromosome Plasmid Recombinant DNA (plasmid) 2 Gene of interest Plasmid put into bacterial cell DNA of chromosome Recombinant bacterium 3 Host cell grown in culture to form a clone of cells containing the cloned gene of interest Gene of Interest Protein expressed by gene of interest Copies of gene Protein harvested Basic research on gene 4 Basic research and various applications Basic research on protein Gene for pest resistance inserted into plants Gene used to alter bacteria for cleaning up toxic waste Protein dissolves blood clots in heart attack therapy Human growth hormone treats stunted growth
7 Restriction Enzymes Restriction enzymes enzymes that cut DNA molecules at a limited number of specific locations based on sequence. Restriction enzymes are able to recognize restriction sites, which are short specific sequences of DNA. Restriction enzymes can cut both DNA strands within the restriction site, most of which are symmetrical The pieces of DNA produced are known as restriction fragments Methylation of adenines or cytosines can protect bacterial DNA from the restriction enzymes
8 Making Recombinant DNA Restriction enzymes cleave the sugar phosphate backbone in both strands of DNA, usually leaving a uneven end. This uneven end produces one longer single-stranded end, which is known as a sticky end. This can be hydrogen bonded with complementary single-stranded DNA. Ligase can then catalyze the formation of new bonds in the sugar phosphate backbone. This method can be used to insert DNA into an existing molecule, forming recombinant DNA.
9 Restriction site DNA Restriction enzyme cuts sugar-phosphate backbones. Sticky end 2 DNA fragment added from another molecule cut by same enzyme. Base pairing occurs. 3 DNA ligase seals strands. One possible combination Recombinant DNA molecule
10 Cloning Genes in Bacterial Plasmids Cloning of a hummingbird gene: 1. Isolate bacterial plasmid and isolate hummingbird DNA. Bacterial plasmid contains the genes amp R and lacz. 2. Both samples of DNA are then digested with the same restriction enzyme. The restriction enzyme chosen only has one site in the bacterial plasmid which is within the lacz gene. 3. The fragments generated will then base pair their complementary sticky ends and DNA ligase can bond the sugar phosphate backbones.
11 Cloning Genes in Bacterial Plasmids Cloning continued 4. The DNA mixture is then added to a bacterial culture with bacteria that have a mutation in their lacz gene. 5. Bacteria is then plated on nutrient agar with ampicillin and X-gal, which will select for colonies that have taken in recombinant plasmid. 6. White colonies that grow on this medium will have taken in recombinant plasmid. The recombinant plasmid will contain different fragments of hummingbird DNA.
12 Selecting for Recombinant Bacteria The medium contains ampicillin and X-gal to identify recombinant bacteria The amp R gene provides ampicillin resistance. Therefore, only bacteria with the amp R gene will survive on medium with ampicillin. This verifies the presence of the plasmid in the bacteria. Only bacteria with functioning β-galactosidase will produce a blue colonies due to cleavage of X-gal by β-galactosidase, which produces a blue product. Therefore, only white colonies will have a recombinant plasmid within (hummingbird DNA will have inserted into the lacz gene of the plasmid).
13 TECHNIQUE Bacterial cell lacz gene Hummingbird cell amp R gene Bacterial plasmid Restriction site Sticky ends Gene of interest Hummingbird DNA fragments Nonrecombinant plasmid Recombinant plasmids Bacteria carrying plasmids RESULTS Colony carrying nonrecombinant plasmid with intact lacz gene Colony carrying recombinant plasmid with disrupted lacz gene One of many bacterial clones
14 Storing Cloned Genes in DNA Libraries The cloning procedure produces colonies of recombinant bacteria that contain foreign genomic DNA. A genomic library is the complete set of plasmidcontaining clones, each carrying copies of a particular segment from the initial genome. A genomic library that is made using bacteria is the collection of recombinant vector clones. A genomic library that is made using bacteriophages is stored as a collection of phage clones. A bacterial artificial chromosome (BAC) is a large plasmid that has been trimmed down and can carry a large DNA insert. This minimizes the number of clones needed.
15 or Recombinant phage DNA Foreign genome cut up with restriction enzyme Large plasmid Large insert with many genes BAC clone Bacterial clones Recombinant plasmids Phage clones (a) Plasmid library (b) Phage library (c) A library of bacterial artificial chromosome (BAC) clones
16 Storing Cloned Genes in DNA Libraries A complementary DNA (cdna) library is made by cloning DNA made in vitro by reverse transcription of all the mrna produced by a particular cell. To do this, mrna is extracted from cells and reverse transcribed to produce DNA that is complementary to the mrna. A cdna library represents only part of the genome only the subset of genes transcribed into mrna in the original cells.
17 DNA in nucleus mrnas in cytoplasm mrna Reverse transcriptase Poly-A tail Degraded mrna DNA strand Primer DNA polymerase cdna
18 Screening a Library for Clones Carrying a Gene of Interest Once clones have be cultured that contain the foreign DNA, clones carrying the gene of interest must be identified. A clone carrying the gene of interest can be identified with a nucleic acid probe having a sequence complementary to the gene. The nucleic acid probe is usually labelled somehow (radioactivity, luminescence, etc.) so that its presence can be identified. This process is called nucleic acid hybridization.
19 Screening a Library for Clones Carrying a Gene of Interest A probe can be synthesized that is complementary to the gene of interest. For example, if the desired gene is 5 GGCT AACT T AGC Then we would synthesize this probe 3 CCGAT T GAAT CG The DNA probe can be used to screen a large number of clones simultaneously for the gene of interest through complementary base pairing. Once identified, the clone carrying the gene of interest can be cultured to amplify the gene of interest. 3 5
20 TECHNIQUE Multiwell plates holding library clones Radioactively labeled probe molecules Probe DNA Gene of interest Single-stranded DNA from cell Film Nylon membrane Nylon Location of membrane DNA with the complementary sequence
21 Bacterial Expression Systems After a gene has been cloned, its protein product can be produced in larger amounts for research. Cloned genes can be expressed as protein in either bacterial or eukaryotic cells. However, due to differences between bacterial and eukaryotic gene expression, several obstacles must be overcome. To overcome differences in promoters and other DNA control sequences, scientists usually employ an expression vector, a cloning vector that contains a highly active prokaryotic promoter upstream of the restriction site for insertion of the eukaryotic gene. To overcome the lack of RNA-splicing, a cdna form of a gene can be used.
22 Eukaryotic Cloning and Expression Systems The use of cultured eukaryotic cells as host cells and yeast artificial chromosomes (YACs) as vectors helps avoid gene expression problems. YACs behave normally in mitosis and can carry more DNA than a plasmid. Eukaryotic hosts can provide the post-translational modifications that many proteins require. One method of introducing recombinant DNA into eukaryotic cells is electroporation, applying a brief electrical pulse to create temporary holes in plasma membranes. Alternatively, scientists can inject DNA into cells using microscopically thin needles. Once inside the cell, the DNA can be incorporated into the cell s DNA by genetic recombination.
23 Amplifying DNA in Vitro: The Polymerase Chain Reaction (PCR) While DNA cloning in cells is effective for cloning large quantities of a gene, other methods can also be used to amplify DNA. The polymerase chain reaction, PCR, can produce many copies of a specific target segment of DNA without the use of a cell. A three-step cycle heating, cooling, and replication brings about a chain reaction that produces an exponentially growing population of identical DNA molecules. The DNA polymerase used is stable at high temperatures.
24 TECHNIQUE 5 3 Target sequence Genomic DNA Denaturation Annealing 3 5 Cycle 1 yields 2 molecules 3 Extension Primers New nucleotides
25 Cycle 2 yields 4 molecules Cycle 3 yields 8 molecules; 2 molecules (in white boxes) match target sequence
26 Concept 20.2 DNA technology allows us to study the sequence, expression, and function of a gene DNA cloning allows researchers to compare genes and alleles between individuals Locate gene expression in a body Determine the role of a gene in an organism Several techniques can be used to analyze the samples obtained from DNA cloning.
27 Gel Electrophoresis One indirect method of rapidly analyzing and comparing genomes is gel electrophoresis. This technique uses a gel as a molecular sieve to separate nucleic acids or proteins by size. A current is applied that causes charged molecules to move through the gel (nucleic acids carry a negative charge on their phosphate groups). Molecules are sorted into bands by their size. Larger molecules generally migrate slower through the gel than smaller molecules. A ladder containing fragments of known sizes can be used to estimate sizes of the fragments.
28 TECHNIQUE Mixture of DNA molecules of different sizes Power source Cathode Anode + 1 Gel Longer molecules Power source + 2 Shorter molecules
29 RESULTS
30 Restriction Fragment Analysis In restriction fragment analysis, DNA fragments produced by restriction enzyme digestion of a DNA molecule are sorted by gel electrophoresis. Restriction fragment analysis is useful for comparing two different DNA molecules, such as two alleles for a gene. The procedure is also used to prepare pure samples of individual fragments which can be isolated and recovered from the gel. However, larger DNA molecules will result in a smear of different sized fragments of DNA, rather than distinct bands.
31 Normal -globin allele Normal allele Sickle-cell allele 175 bp 201 bp Large fragment DdeI DdeI DdeI DdeI Large fragment Sickle-cell mutant -globin allele 376 bp Large fragment 201 bp 175 bp 376 bp DdeI DdeI DdeI (a) DdeI restriction sites in normal and sickle-cell alleles of -globin gene (b) Electrophoresis of restriction fragments from normal and sickle-cell alleles
32 Southern Blotting A technique called Southern blotting combines gel electrophoresis of DNA fragments with nucleic acid hybridization. Specific DNA fragments can be identified by Southern blotting, using labeled probes that hybridize to the DNA immobilized on a blot of the gel on a nitrocellulose membrane. This is useful when too many bands are produced and the sequence of interest cannot be resolved from the other bands in a gel.
33 TECHNIQUE DNA + restriction enzyme Restriction fragments I II III Nitrocellulose membrane (blot) Heavy weight Gel Sponge I Normal -globin allele II Sickle-cell allele III Heterozygote Alkaline solution Paper towels 1 Preparation of restriction fragments 2 Gel electrophoresis 3 DNA transfer (blotting) Radioactively labeled probe for -globin gene I II III Probe base-pairs with fragments I II III Nitrocellulose blot Fragment from sickle-cell -globin allele Fragment from normal -globin allele 4 Hybridization with radioactive probe 5 Probe detection Film over blot
34 DNA Sequencing Once a fragment of DNA is cloned, its sequence can be determined through the dideoxyribonucleotide chain termination method, developed by Frederick Sanger. Modified nucleotides called dideoxyribonucleotides (ddntp) attach to synthesized DNA strands of different lengths. Each type of ddntp is tagged with a distinct fluorescent label that identifies the nucleotide at the end of each DNA fragment. The DNA sequence can be read from the resulting spectrogram.
35 TECHNIQUE DNA (template strand) Primer Deoxyribonucleotides Dideoxyribonucleotides (fluorescently tagged) DNA polymerase datp dctp dttp dgtp ddatp ddctp ddttp ddgtp
36 TECHNIQUE DNA (template strand) Labeled strands Shortest Longest
37 TECHNIQUE Direction of movement of strands Longest labeled strand Detector RESULTS Laser Last base of longest labeled strand Shortest labeled strand Last base of shortest labeled strand
38 Studying the Expression of Single Genes Nucleic acid probes can hybridize with mrnas transcribed from a gene These probes can be used to identify where or when a gene is transcribed in an organism For example, changes in the expression of a gene during embryonic development can be tested using: Northern blotting Reverse transcriptase-polymerase chain reaction Both methods are used to compare mrna from different developmental stages
39 Northern Blotting Northern blotting combines gel electrophoresis of mrna (instead of DNA like in the Southern blot) followed by hybridization with a probe on a membrane. Detection of a hybridized probe would indicate the presence of mrna with that particular sequence in that sample. Identification of mrna at a particular developmental stage suggests that possibly protein functions at that stage as well.
40 RT-PCR Reverse transcriptase-polymerase chain reaction (RT-PCR) is quicker and more sensitive than a Northern blot. Reverse transcriptase is added to mrna to make cdna, which serves as a template for PCR amplification of the gene of interest. The products are run on a gel and the mrna of interest identified and quantified in different samples.
41 TECHNIQUE 1 cdna synthesis mrnas 2 PCR amplification Primers cdnas 3 Gel electrophoresis -globin gene RESULTS Embryonic stages
42 In situ hybridization In situ hybridization uses fluorescent dyes attached to probes to identify the location of specific mrnas in place in the intact organism 50 µm
43 Studying the Expression of Interacting Groups of Genes Now that entire genomes of various organisms have been sequenced, it is possible to look at expression of large groups of genes to determine which genes are transcribed in different situations. Groups of genes that are expressed coordinately can also be found. Automation has allowed scientists to measure expression of thousands of genes at one time using DNA microarray assays DNA microarray assays compare patterns of gene expression in different tissues, at different times, or under different conditions
44 TECHNIQUE 1 Isolate mrna. Tissue sample 2 Make cdna by reverse transcription, using fluorescently labeled nucleotides. mrna molecules 3 Apply the cdna mixture to a microarray, a different gene in each spot. The cdna hybridizes with any complementary DNA on the microarray. Labeled cdna molecules (single strands) DNA fragments representing specific genes DNA microarray 4 Rinse off excess cdna; scan microarray for fluorescence. Each fluorescent spot represents a gene expressed in the tissue sample. DNA microarray with 2,400 human genes
45
46 Determining Gene Function One way to determine function is to disable the gene and observe the consequences. Using in vitro mutagenesis, specific mutations are introduced into a cloned gene, to knock out or disable the gene. When the mutated gene is returned to the cell, the normal gene s function might be determined by examining the mutant s phenotype Gene expression can also be silenced using RNA interference (RNAi) sirnas can be used to break down or block the gene s mrna
Biotechnology. Chapter 20. Biology Eighth Edition Neil Campbell and Jane Reece. PowerPoint Lecture Presentations for
Chapter 20 Biotechnology PowerPoint Lecture Presentations for Biology Eighth Edition Neil Campbell and Jane Reece Lectures by Chris Romero, updated by Erin Barley with contributions from Joan Sharp Copyright
More informationOverview: The DNA Toolbox
Overview: The DNA Toolbox Sequencing of the genomes of more than 7,000 species was under way in 2010 DNA sequencing has depended on advances in technology, starting with making recombinant DNA In recombinant
More informationRecombinant DNA recombinant DNA DNA cloning gene cloning
DNA Technology Recombinant DNA In recombinant DNA, DNA from two different sources, often two species, are combined into the same DNA molecule. DNA cloning permits production of multiple copies of a specific
More informationCHAPTER 20 DNA TECHNOLOGY AND GENOMICS. Section A: DNA Cloning
Section A: DNA Cloning 1. DNA technology makes it possible to clone genes for basic research and commercial applications: an overview 2. Restriction enzymes are used to make recombinant DNA 3. Genes can
More informationBiotechnology. Chapter 20. Biology Eighth Edition Neil Campbell and Jane Reece. PowerPoint Lecture Presentations for
Chapter 20 Biotechnology PowerPoint Lecture Presentations for Biology Eighth Edition Neil Campbell and Jane Reece Lectures by Chris Romero, updated by Erin Barley with contributions from Joan Sharp Copyright
More informationOverview: The DNA Toolbox
Overview: The DNA Toolbox Sequencing of the genomes of more than 7,000 species was under way in 2010 DNA sequencing has depended on advances in technology, starting with making recombinant DNA In recombinant
More informationBiotechnology. Chapter 20. Biology Eighth Edition Neil Campbell and Jane Reece. PowerPoint Lecture Presentations for
The image cannot be displayed. Your computer may not have enough memory to open the image, or the image may have been corrupted. Restart your computer, and then open the file again. If the red x still
More informationBiotechnology DNA technology
Biotechnology Biotechnology is the manipulation of organisms or their components to make useful products The applications of DNA technology affect everything from agriculture, to criminal law, to medical
More informationChapter 20 Recombinant DNA Technology. Copyright 2009 Pearson Education, Inc.
Chapter 20 Recombinant DNA Technology Copyright 2009 Pearson Education, Inc. 20.1 Recombinant DNA Technology Began with Two Key Tools: Restriction Enzymes and DNA Cloning Vectors Recombinant DNA refers
More informationChapter 20 DNA Technology & Genomics. If we can, should we?
Chapter 20 DNA Technology & Genomics If we can, should we? Biotechnology Genetic manipulation of organisms or their components to make useful products Humans have been doing this for 1,000s of years plant
More informationMolecular Genetics Techniques. BIT 220 Chapter 20
Molecular Genetics Techniques BIT 220 Chapter 20 What is Cloning? Recombinant DNA technologies 1. Producing Recombinant DNA molecule Incorporate gene of interest into plasmid (cloning vector) 2. Recombinant
More information2014 Pearson Education, Inc. CH 8: Recombinant DNA Technology
CH 8: Recombinant DNA Technology Biotechnology the use of microorganisms to make practical products Recombinant DNA = DNA from 2 different sources What is Recombinant DNA Technology? modifying genomes
More informationCH 8: Recombinant DNA Technology
CH 8: Recombinant DNA Technology Biotechnology the use of microorganisms to make practical products Recombinant DNA = DNA from 2 different sources What is Recombinant DNA Technology? modifying genomes
More informationChapter 20: Biotechnology
Chapter 20: Biotechnology 1. DNA Sequencing 2. DNA Cloning 3. Studying Gene Expression 4. Manipulating Genomes 5. herapeutic & Diagnostic echniques 1. DNA Sequencing Chapter Reading pp. 409-412 DNA Sequencing
More informationChapter 6 - Molecular Genetic Techniques
Chapter 6 - Molecular Genetic Techniques Two objects of molecular & genetic technologies For analysis For generation Molecular genetic technologies! For analysis DNA gel electrophoresis Southern blotting
More informationRecombinant DNA Technology. The Role of Recombinant DNA Technology in Biotechnology. yeast. Biotechnology. Recombinant DNA technology.
PowerPoint Lecture Presentations prepared by Mindy Miller-Kittrell, North Carolina State University C H A P T E R 8 Recombinant DNA Technology The Role of Recombinant DNA Technology in Biotechnology Biotechnology?
More informationAP Biology Day 34. Monday, November 14, 2016
AP Biology Day 34 Monday, November 14, 2016 Essen%al knowledge standards 3.A.1: DNA, and in some cases RNA, is the primary source of heritable informa%on 3.A.1.e: Gene%c engineering techniques can manipulate
More information7.1 Techniques for Producing and Analyzing DNA. SBI4U Ms. Ho-Lau
7.1 Techniques for Producing and Analyzing DNA SBI4U Ms. Ho-Lau What is Biotechnology? From Merriam-Webster: the manipulation of living organisms or their components to produce useful usually commercial
More informationLecture Four. Molecular Approaches I: Nucleic Acids
Lecture Four. Molecular Approaches I: Nucleic Acids I. Recombinant DNA and Gene Cloning Recombinant DNA is DNA that has been created artificially. DNA from two or more sources is incorporated into a single
More informationBiotechnolog y and DNA Technology
PowerPoint Lecture Presentations prepared by Bradley W. Christian, McLennan Community College C H A P T E R 9 Biotechnolog y and DNA Technology Introduction to Biotechnology Biotechnology: the use of microorganisms,
More informationBIOLOGY - CLUTCH CH.20 - BIOTECHNOLOGY.
!! www.clutchprep.com CONCEPT: DNA CLONING DNA cloning is a technique that inserts a foreign gene into a living host to replicate the gene and produce gene products. Transformation the process by which
More informationBIOTECHNOLOGY. Sticky & blunt ends. Restriction endonucleases. Gene cloning an overview. DNA isolation & restriction
BIOTECHNOLOGY RECOMBINANT DNA TECHNOLOGY Recombinant DNA technology involves sticking together bits of DNA from different sources. Made possible because DNA & the genetic code are universal. 2004 Biology
More informationDNA Technology and Genomics
Chapter 20 DNA echnology and enomics PowerPoint Lectures for Biology, Seventh Edition Neil Campbell and Jane Reece Lectures by Chris Romero Overview: Understanding and Manipulating enomes One of the greatest
More informationMolecular Cell Biology - Problem Drill 11: Recombinant DNA
Molecular Cell Biology - Problem Drill 11: Recombinant DNA Question No. 1 of 10 1. Which of the following statements about the sources of DNA used for molecular cloning is correct? Question #1 (A) cdna
More informationB. Incorrect! Ligation is also a necessary step for cloning.
Genetics - Problem Drill 15: The Techniques in Molecular Genetics No. 1 of 10 1. Which of the following is not part of the normal process of cloning recombinant DNA in bacteria? (A) Restriction endonuclease
More informationChapter 20: Biotechnology
Name Period The AP Biology exam has reached into this chapter for essay questions on a regular basis over the past 15 years. Student responses show that biotechnology is a difficult topic. This chapter
More informationCHAPTER 9 DNA Technologies
CHAPTER 9 DNA Technologies Recombinant DNA Artificially created DNA that combines sequences that do not occur together in the nature Basis of much of the modern molecular biology Molecular cloning of genes
More informationXXII DNA cloning and sequencing. Outline
XXII DNA cloning and sequencing 1) Deriving DNA for cloning Outline 2) Vectors; forming recombinant DNA; cloning DNA; and screening for clones containing recombinant DNA [replica plating and autoradiography;
More informationChapter 10 Genetic Engineering: A Revolution in Molecular Biology
Chapter 10 Genetic Engineering: A Revolution in Molecular Biology Genetic Engineering Direct, deliberate modification of an organism s genome bioengineering Biotechnology use of an organism s biochemical
More informationSelected Techniques Part I
1 Selected Techniques Part I Gel Electrophoresis Can be both qualitative and quantitative Qualitative About what size is the fragment? How many fragments are present? Is there in insert or not? Quantitative
More informationDNA Technology. B. Using Bacteria to Clone Genes: Overview:
DNA Technology A. Basic Vocabulary: is DNA from 2 different sources that is combined. is the direct manipulation of genes for practical purposes. literally means or in a test tube or flask. is the manipulation
More informationRecitation CHAPTER 9 DNA Technologies
Recitation CHAPTER 9 DNA Technologies DNA Cloning: General Scheme A cloning vector and eukaryotic chromosomes are separately cleaved with the same restriction endonuclease. (A single chromosome is shown
More informationDNA Technology and Genomics
DNA Technology and Genomics I. DNA cloning permits production of many copies of a specific gene or other DNA segment. A. To study a particular gene, scientists needed to develop methods to isolate the
More informationChapter 8: Recombinant DNA. Ways this technology touches us. Overview. Genetic Engineering
Chapter 8 Recombinant DNA and Genetic Engineering Genetic manipulation Ways this technology touches us Criminal justice The Justice Project, started by law students to advocate for DNA testing of Death
More informationGenetic Engineering & Recombinant DNA
Genetic Engineering & Recombinant DNA Chapter 10 Copyright The McGraw-Hill Companies, Inc) Permission required for reproduction or display. Applications of Genetic Engineering Basic science vs. Applied
More informationBiotechnology and DNA Technology
11/27/2017 PowerPoint Lecture Presentations prepared by Bradley W. Christian, McLennan Community College CHAPTER 9 Biotechnology and DNA Technology Introduction to Biotechnology Learning Objectives Compare
More informationAP Biology Gene Expression/Biotechnology REVIEW
AP Biology Gene Expression/Biotechnology REVIEW Multiple Choice Identify the choice that best completes the statement or answers the question. 1. Gene expression can be a. regulated before transcription.
More informationChapter 9 Genetic Engineering
Chapter 9 Genetic Engineering Biotechnology: use of microbes to make a protein product Recombinant DNA Technology: Insertion or modification of genes to produce desired proteins Genetic engineering: manipulation
More informationGenetics and Genomics in Medicine Chapter 3. Questions & Answers
Genetics and Genomics in Medicine Chapter 3 Multiple Choice Questions Questions & Answers Question 3.1 Which of the following statements, if any, is false? a) Amplifying DNA means making many identical
More information-Is the process of manipulating genes and genomes
Genetic Engineering -Is the process of manipulating genes and genomes Biotechnology -Is the process of manipulating organisms or their components for the purpose of making useful products Restriction Enzymes
More informationBiotechnology Chapter 20
Biotechnology Chapter 20 DNA Cloning DNA Cloning AKA Plasmid-based transformation or molecular cloning First off-let s sum up what happens. A plasmid is taken from a bacteria A gene is inserted into the
More information2054, Chap. 14, page 1
2054, Chap. 14, page 1 I. Recombinant DNA technology (Chapter 14) A. recombinant DNA technology = collection of methods used to perform genetic engineering 1. genetic engineering = deliberate modification
More informationMultiple choice questions (numbers in brackets indicate the number of correct answers)
1 Multiple choice questions (numbers in brackets indicate the number of correct answers) February 1, 2013 1. Ribose is found in Nucleic acids Proteins Lipids RNA DNA (2) 2. Most RNA in cells is transfer
More informationSTANDARD CLONING PROCEDURES. Shotgun cloning (using a plasmid vector and E coli as a host).
STANDARD CLONING PROCEDURES Shotgun cloning (using a plasmid vector and E coli as a host). 1) Digest donor DNA and plasmid DNA with the same restriction endonuclease 2) Mix the fragments together and treat
More informationBootcamp: Molecular Biology Techniques and Interpretation
Bootcamp: Molecular Biology Techniques and Interpretation Bi8 Winter 2016 Today s outline Detecting and quantifying nucleic acids and proteins: Basic nucleic acid properties Hybridization PCR and Designing
More informationMotivation From Protein to Gene
MOLECULAR BIOLOGY 2003-4 Topic B Recombinant DNA -principles and tools Construct a library - what for, how Major techniques +principles Bioinformatics - in brief Chapter 7 (MCB) 1 Motivation From Protein
More informationBiotechnology. Biotechnology is difficult to define but in general it s the use of biological systems to solve problems.
MITE 2 S Biology Biotechnology Summer 2004 Austin Che Biotechnology is difficult to define but in general it s the use of biological systems to solve problems. Recombinant DNA consists of DNA assembled
More informationMolecular Cloning. Genomic DNA Library: Contains DNA fragments that represent an entire genome. cdna Library:
Molecular Cloning Genomic DNA Library: Contains DNA fragments that represent an entire genome. cdna Library: Made from mrna, and represents only protein-coding genes expressed by a cell at a given time.
More informationPLNT2530 (2018) Unit 6b Sequence Libraries
PLNT2530 (2018) Unit 6b Sequence Libraries Molecular Biotechnology (Ch 4) Analysis of Genes and Genomes (Ch 5) Unless otherwise cited or referenced, all content of this presenataion is licensed under the
More informationChapter 13: Biotechnology
Chapter Review 1. Explain why the brewing of beer is considered to be biotechnology. The United Nations defines biotechnology as any technological application that uses biological system, living organism,
More information4/26/2015. Cut DNA either: Cut DNA either:
Ch.20 Enzymes that cut DNA at specific sequences (restriction sites) resulting in segments of DNA (restriction fragments) Typically 4-8 bp in length & often palindromic Isolated from bacteria (Hundreds
More informationBiotechnology: DNA Technology & Genomics
Chapter 20. Biotechnology: DNA Technology & Genomics 2003-2004 1 The BIG Questions! How can we use our knowledge of DNA to: " diagnose disease or defect? " cure disease or defect? " change/improve organisms?!
More informationCHEM 4420 Exam I Spring 2013 Page 1 of 6
CHEM 4420 Exam I Spring 2013 Page 1 of 6 Name Use complete sentences when requested. There are 100 possible points on this exam. The multiple choice questions are worth 2 points each. All other questions
More informationVirus- infectious particle consisting of nucleic acid packaged in a protein coat.
Chapter 19 Virus- infectious particle consisting of nucleic acid packaged in a protein coat. Most scientists consider viruses non-living because they cannot reproduce or carry out metabolic activities
More informationAGRO/ANSC/BIOL/GENE/HORT 305 Fall, 2017 Recombinant DNA Technology (Chpt 20, Genetics by Brooker) Lecture outline: (#14)
AGRO/ANSC/BIOL/GENE/HORT 305 Fall, 2017 Recombinant DNA Technology (Chpt 20, Genetics by Brooker) Lecture outline: (#14) - RECOMBINANT DNA TECHNOLOGY is the use of in vitro molecular techniques to isolate
More informationAP Biology. Chapter 20. Biotechnology: DNA Technology & Genomics. Biotechnology. The BIG Questions. Evolution & breeding of food plants
What do you notice about these phrases? radar racecar Madam I m Adam Able was I ere I saw Elba a man, a plan, a canal, Panama Was it a bar or a bat I saw? Chapter 20. Biotechnology: DNA Technology & enomics
More informationRestriction Enzymes (endonucleases)
In order to understand and eventually manipulate DNA (human or otherwise) an array of DNA technologies have been developed. Here are some of the tools: Restriction Enzymes (endonucleases) In order to manipulate
More informationThe Biotechnology Toolbox
Chapter 15 The Biotechnology Toolbox Cutting and Pasting DNA Cutting DNA Restriction endonuclease or restriction enzymes Cellular protection mechanism for infected foreign DNA Recognition and cutting specific
More informationCh.15 Section 4 Regulation of Gene Expression pgs Complete the attached Active Reading Guides for the above sections.
AP Biology 2018-2019 Summer Assignment Due Wednesday 9/5/2018 Text Book Reading Ch.13 The Molecular Basis of Life pgs. 245-267 Ch.15 Section 4 Regulation of Gene Expression pgs. 307-309 Active Reading
More informationGenetic Fingerprinting
Genetic Fingerprinting Introduction DA fingerprinting In the R & D sector: -involved mostly in helping to identify inherited disorders. In forensics: -identification of possible suspects involved in offences.
More informationComputational Biology I LSM5191
Computational Biology I LSM5191 Lecture 5 Notes: Genetic manipulation & Molecular Biology techniques Broad Overview of: Enzymatic tools in Molecular Biology Gel electrophoresis Restriction mapping DNA
More informationRestriction Enzymes (Site-Specific Endonuclease) Enzymes that recognize and cleave dsdna in a highly sequence specific manner.
Enzymes Restriction Enzymes (Site-Specific Endonuclease) Enzymes that recognize and cleave dsdna in a highly sequence specific manner. Generally recognize an inverted repeat sequence 4, 6, or 8 base pairs
More informationCHAPTER 08: RECOMBINANT DNA TECHNOLOGY Pearson Education, Inc.
CHAPTER 08: RECOMBINANT DNA TECHNOLOGY The Role of Recombinant DNA Technology in Biotechnology Biotechnology the use of microorganisms to make practical products Recombinant DNA technology Intentionally
More informationChapter 20: Biotechnology. 1. DNA Sequencing 3/26/2017. DNA Sequencing. 2. DNA Cloning. 3. Studying Gene Expression. 4. Manipulating Genomes
hapter 0: Biotechnology 1. DN Sequencing. DN loning 3. Studying ene Expression 4. Manipulating enomes 5. herapeutic & Diagnostic echniques 1. DN Sequencing hapter Reading pp. 409-41 EHNIQUE DN (template
More informationUnit 8: Genomics Guided Reading Questions (150 pts total)
Name: AP Biology Biology, Campbell and Reece, 7th Edition Adapted from chapter reading guides originally created by Lynn Miriello Chapter 18 The Genetics of Viruses and Bacteria Unit 8: Genomics Guided
More informationGENETICS EXAM 3 FALL a) is a technique that allows you to separate nucleic acids (DNA or RNA) by size.
Student Name: All questions are worth 5 pts. each. GENETICS EXAM 3 FALL 2004 1. a) is a technique that allows you to separate nucleic acids (DNA or RNA) by size. b) Name one of the materials (of the two
More informationFatchiyah
Fatchiyah Email: fatchiya@yahoo.co.id RNAs: mrna trna rrna RNAi DNAs: Protein: genome DNA cdna mikro-makro mono-poly single-multi Analysis: Identification human and animal disease Finger printing Sexing
More informationA Lot of Cutting and Pasting Going on Here Recombinant DNA and Biotechnology
A Lot of Cutting and Pasting Going on Here Recombinant DNA and Biotechnology How Are Large DNA Molecules Analyzed? Naturally occurring enzymes that cleave and repair DNA are used in the laboratory to manipulate
More informationBiotechnology DNA cloning yields multiple copies of a gene or other DNA segment
20 Biotechnology KEY CONCEPS 20.1 DNA cloning yields multiple copies of a gene or other DNA segment 20.2 DNA technology allows us to study the sequence, expression, and function of a gene 20.3 Cloning
More informationCELL BIOLOGY - CLUTCH CH TECHNIQUES IN CELL BIOLOGY.
!! www.clutchprep.com CONCEPT: LIGHT MICROSCOPE A light microscope uses light waves and magnification to view specimens Can be used to visualize transparent, and some cellular components - Can visualize
More informationDesign. Construction. Characterization
Design Construction Characterization DNA mrna (messenger) A C C transcription translation C A C protein His A T G C T A C G Plasmids replicon copy number incompatibility selection marker origin of replication
More informationBio 101 Sample questions: Chapter 10
Bio 101 Sample questions: Chapter 10 1. Which of the following is NOT needed for DNA replication? A. nucleotides B. ribosomes C. Enzymes (like polymerases) D. DNA E. all of the above are needed 2 The information
More informationLearning Objectives :
Learning Objectives : Understand the basic differences between genomic and cdna libraries Understand how genomic libraries are constructed Understand the purpose for having overlapping DNA fragments in
More informationDESIGNER GENES - BIOTECHNOLOGY
DESIGNER GENES - BIOTECHNOLOGY Technology to manipulate DNA techniques often called genetic engineering or Recombinant DNA Technology-Technology used to manipulate DNA Procedures often called genetic engineering
More informationBi 8 Lecture 4. Ellen Rothenberg 14 January Reading: from Alberts Ch. 8
Bi 8 Lecture 4 DNA approaches: How we know what we know Ellen Rothenberg 14 January 2016 Reading: from Alberts Ch. 8 Central concept: DNA or RNA polymer length as an identifying feature RNA has intrinsically
More information3. Translation. 2. Transcription. 1. Replication. and functioning through their expression in. Genes are units perpetuating themselves
Central Dogma Genes are units perpetuating themselves and functioning through their expression in the form of proteins 1 DNA RNA Protein 2 3 1. Replication 2. Transcription 3. Translation Spring 2002 21
More informationUnit 2: Metabolism and Survival Sub-Topic (2.7) Genetic Control of Metabolism (2.8) Ethical considerations in the use of microorganisms
Unit 2: Metabolism and Survival Sub-Topic (2.7) Genetic Control of Metabolism (2.8) Ethical considerations in the use of microorganisms Duncanrig Secondary JHM&MHC 2015 Page 1 of 18 On completion of this
More informationGenetic Fingerprinting
Genetic Fingerprinting Introduction DA fingerprinting In the R & D sector: -involved mostly in helping to identify inherited disorders. In forensics: -identification of possible suspects involved in offences.
More informationBiotechnology. DNA Cloning Finding Needles in Haystacks. DNA Sequencing. Genetic Engineering. Gene Therapy
Biotechnology DNA Cloning Finding Needles in Haystacks DNA Sequencing Genetic Engineering Gene Therapy What is DNA Cloning? Set of methods that uses live cells to make many identical copies of a DNA fragment
More informationChapter 15 Recombinant DNA and Genetic Engineering. Restriction Enzymes Function as Nature s Pinking Shears
Chapter 15 Recombinant DNA and Genetic Engineering In this chapter you will learn How restriction enzyme work and why they are essential to DNA technology. About various procedures such as cloning and
More informationChapter 4. Recombinant DNA Technology
Chapter 4 Recombinant DNA Technology 5. Plasmid Cloning Vectors Plasmid Plasmids Self replicating Double-stranded Mostly circular DNA ( 500 kb) Linear : Streptomyces, Borrelia burgdorferi Replicon
More informationCh.15 Section 4 Regulation of Gene Expression pgs Complete the attached Active Reading Guides for the above sections.
AP Biology 2018-2019 Summer Assignment Due Wednesday 9/5/2018 Text Book Reading Ch.13 The Molecular Basis of Life pgs. 245-267 Ch.15 Section 4 Regulation of Gene Expression pgs. 307-309 Active Reading
More informationDNA and Biotechnology Form of DNA Form of DNA Form of DNA Form of DNA Replication of DNA Replication of DNA
21 DNA and Biotechnology DNA and Biotechnology OUTLINE: Replication of DNA Gene Expression Mutations Regulating Gene Activity Genetic Engineering Genomics DNA (deoxyribonucleic acid) Double-stranded molecule
More informationBIOTECHNOLOGY. Biotechnology is the process by which living organisms are used to create new products THE ORGANISMS
BIOTECHNOLOGY Biotechnology is the process by which living organisms are used to create new products THE ORGANISMS Bacteria: are prokaryotic organisms that contain circular DNA and no organelles. They
More informationApplicazioni biotecnologiche
Applicazioni biotecnologiche Analisi forense Sintesi di proteine ricombinanti Restriction Fragment Length Polymorphism (RFLP) Polymorphism (more fully genetic polymorphism) refers to the simultaneous occurrence
More informationGenetics Lecture 21 Recombinant DNA
Genetics Lecture 21 Recombinant DNA Recombinant DNA In 1971, a paper published by Kathleen Danna and Daniel Nathans marked the beginning of the recombinant DNA era. The paper described the isolation of
More informationLearning Objectives. 2. Restriction Endonucleases 3. Cloning 4. Genetic Engineering 5. DNA libraries 6. PCR 7. DNA Fingerprinting
Fig. 13-CO, p.330 Learning Objectives 1. Purification & detection of nucleic acids. 2. Restriction Endonucleases 3. Cloning 4. Genetic Engineering 5. DNA libraries 6. PCR 7. DNA Fingerprinting Gel Electrophoresis
More informationChapter 8 Recombinant DNA Technology. 10/1/ MDufilho
Chapter 8 Recombinant DNA Technology 10/1/2017 1 MDufilho The Role of Recombinant DNA Technology in Biotechnology Biotechnology? Recombinant deoxyribonucleic acid (DNA) technology Intentionally modifying
More informationBasic lab techniques
Basic lab techniques Sandrine Dudoit Bioconductor short course Summer 2002 Copyright 2002, all rights reserved Lab techniques Basic lab techniques for nucleic acids Hybridization. Cut: restriction enzymes.
More informationExam 2 Key - Spring 2008 A#: Please see us if you have any questions!
Page 1 of 5 Exam 2 Key - Spring 2008 A#: Please see us if you have any questions! 1. A mutation in which parts of two nonhomologous chromosomes change places is called a(n) A. translocation. B. transition.
More informationMolecular Cloning. Restriction Enzymes and Ligases
Tools in Genetic engineering The science of using living systems to benefit humankind is called biotechnology. Technically speaking, the domestication of plants and animals through farming and breeding
More informationConcept 13.1 Recombinant DNA Can Be Made in the Laboratory
13 Biotechnology Concept 13.1 Recombinant DNA Can Be Made in the Laboratory It is possible to modify organisms with genes from other, distantly related organisms. Recombinant DNA is a DNA molecule made
More informationChapter 9. Biotechnology and DNA Technology
Chapter 9 Biotechnology and DNA Technology SLOs Compare and contrast biotechnology, recombinant DNA technology, and genetic engineering. Identify the roles of a clone and a vector in making recombined
More informationBIOTECHNOLOGY : PRINCIPLES AND PROCESSES
CHAPTER 11 BIOTECHNOLOGY : PRINCIPLES AND PROCESSES POINTS TO REMEMBER Bacteriophage : A virus that infects bacteria. Bioreactor : A large vessel in which raw materials are biologically converted into
More informationRecombinant DNA Technology
Recombinant DNA Technology Common General Cloning Strategy Target DNA from donor organism extracted, cut with restriction endonuclease and ligated into a cloning vector cut with compatible restriction
More informationBiotechnology. Cloning. Transformation 2/4/ glue DNA
Biotechnology Cloning The production of multiple copies of a single gene (gene cloning) For basic research on genes and their protein products To make a protein product (insulin, human growth hormone)
More informationRecombinant DNA Technology
History of recombinant DNA technology Recombinant DNA Technology (DNA cloning) Majid Mojarrad Recombinant DNA technology is one of the recent advances in biotechnology, which was developed by two scientists
More informationLecture 25 (11/15/17)
Lecture 25 (11/15/17) Reading: Ch9; 328-332 Ch25; 990-995, 1005-1012 Problems: Ch9 (study-guide: applying); 1,2 Ch9 (study-guide: facts); 7,8 Ch25 (text); 1-3,5-7,9,10,13-15 Ch25 (study-guide: applying);
More informationDNA Technology. Asilomar Singer, Zinder, Brenner, Berg
DNA Technology Asilomar 1973. Singer, Zinder, Brenner, Berg DNA Technology The following are some of the most important molecular methods we will be using in this course. They will be used, among other
More information