Reading Lecture 3: 24-25, 45, Lecture 4: 66-71, Lecture 3. Vectors. Definition Properties Types. Transformation

Save this PDF as:

Size: px
Start display at page:

Download "Reading Lecture 3: 24-25, 45, Lecture 4: 66-71, Lecture 3. Vectors. Definition Properties Types. Transformation"

Transcription

1 Lecture 3 Reading Lecture 3: 24-25, 45, Lecture 4: 66-71, Vectors Definition Properties Types Transformation 56

2 VECTORS- Definition Vectors are carriers of a DNA fragment of interest Insert Allow the fragment to be replicated in a host cell Means of producing many copies of a specific fragment Bacteria does all the work 57

3 Types of Vectors Plasmids DNA inserts less than 20 kb Phage Inserts in the 20 kb range Cosmids Inserts up to 45 kb BACs DNA fragments up to 300 kb YACs DNA fragments greater than 1 Mb 58

4 Requirements of a Vector Needs a replicon Needs to be able to replicate itself Origin of replication Needs a place to put DNA to be cloned Unique restriction enzyme cleavage site Cloning site Must be selectable 59 Need to be able to distinguish which cells carry the vector Selectable marker gene (antibiotic resistance)

5 Plasmids Extrachromosomal replicating DNA Double-stranded circular DNA Genetically homogenous Stably inherited Can be present in a bacterial cell in multiple copies Not essential for cell s existence Plasmids E. Coli chromosome 60

6 Plasmid Discovery Plasmids are found in bacteria with different phenotypes 3 Classes Virulence plasmids Encode toxin genes Drug-resistance plasmids Confer resistance to antibiotics Conjugation plasmids Encode genes required for bacterial conjugation 61

7 Laboratory Plasmid Characteristics: Origin of Replication Origin of replication Various mechanisms Figs. 4.4 and 4.5 in text F plasmid ori One copy per cell ColEI ori Relaxed replication ~25 copies per cell 62

8 Laboratory Plasmid Characteristics: MCS A polylinker or multiple cloning site (MCS) is an artificially synthesized DNA sequence that is inserted into a vector and carries a set of restriction enzyme cleavage sites unique in the plasmid. The polylinker is integrated into the lacz gene. 63

9 Laboratory Plasmid Characteristics: Selectable Marker- Antibiotic Resistance Ampicillin resistance (Amp r ) Ampicillin inhibits cell wall synthesis Stops bacterial growth Amp r gene product hydrolyzes ampicillin Eliminates ampicillin, cells with plasmid grow 64 Tetracycline resistance (Tet r ) Tetracycline inhibits translation Cell death Tet r gene product prevents tetracycline accumulation Eliminates tetracycline, cells with plasmid grow

10 Transformation Bacterial take-up of DNA Way to introduce your plasmid into an E. coli host Bacteria take up DNA and become transformed Bacteria grow and form colonies Plasmid can replicate and express its genes in the bacterial host Growth of bacteria of selective media will depend on the presence or absence of plasmid 65

11 Selective Media Transformed Bacteria 66 LB LB + Amp

12 Evolution of Plasmid Cloning pbr322 First plasmid cloning vector Ori Few unique restriction enzyme sites Two selectable markers Amp r Tet r Vectors 67

13 More Advanced Plasmid Vectors: puc Developed at UCD by J. Messing puc plasmids carry an ori mutation Increased copy number MCS Polylinker with multiple unique RE sites Amp R and Blue/white selection LacZ gene 68

14 Blue/White Selection for Insert a-complementation system using the E. coli lac operon The lac operon In bacteria, an operon is a cluster of genes Encodes 3 genes LacZ - b-galactosidase LacY- Permease Lac A - Transacetylase Regulated by the lac promoter and repressor The Lac Operon Lacp lacz lacy laca 69

15 The Lac Operon Repressor protein binds to the operator Blocks transcription IPTG can induce expression of the lac operon Lessens repressors affinity for operator 70

16 Product of the lacz gene Activity can be measured in live cells Can cleave the chromogenic substrate X-gal Cleavage of X-gal produces a blue color Can exploit this by using a-complementation Vector expresses N-terminal (a-fragment) of b-gal Host cell expresses C- terminal (w-fragment) Two fragments combine to form functional b-gal 71 b-galactosidase

17 Blue/White Selection in Plasmids DNA for the lac promoter and N- terminus of the lacz gene flank the MCS Transform vector into cells producing C- terminus Plate on media containing IPTG and X- gal 72

18 73 Blue/White Color Selection Vector WITHOUT insert Get blue colonies LacZ gene product is made Cleaves X-gal Get blue precipitate Vector WITH insert Get white colonies Insert disrupts the coding of the lacz gene product No functional b-gal Can t cleave X-gal No blue precipitate

19 pbluescript Commonly used modern plasmid Sold by Stratagene 2 origins or replication ColEI f1 origin Can do ss DNA synthesis T7 and T3 RNA polymerase promoters ss RNA for probes Universal primers 74

20 75 MCS of pbluescript

21 Methods of Transformation Competent cells Chemically treated with CaCl 2 Makes membrane leaky Heat shock to stimulate uptake 76

22 Methods of Transformation Electroporation Uses a short pulse of electrical charge Causes transient opening in cell membrane DNA can get in More efficient than competent cells Used frequently for mammalian cells 77

Prokaryotic Transcription

Prokaryotic Transcription Prokaryotic Transcription Contents 1 The Lactose Intolerance of Bacteria 2 The Lac Operon 3 Lac Operon Simulation 4 LacZ as a reporter gene The Lactose Intolerance of Bacteria The standard growth kinetics

More information

2054, Chap. 14, page 1

2054, 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 information

CHAPTER 9 DNA Technologies

CHAPTER 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 information

Recombinant DNA Technology

Recombinant 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 information

Learning Objectives :

Learning 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 information

Lecture Four. Molecular Approaches I: Nucleic Acids

Lecture 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 information

CHAPTER 20 DNA TECHNOLOGY AND GENOMICS. Section A: DNA Cloning

CHAPTER 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 information

HiPer Plasmid DNA Cloning Teaching Kit

HiPer Plasmid DNA Cloning Teaching Kit HiPer Plasmid DNA Cloning Teaching Kit Product Code: HTBM022 Number of experiments that can be performed: 5 Duration of Experiment: 4 days Day 1- Preparation of media and revival of E. coli Host Day2-

More information

Cloning in bacteria. Presenter: Vito Baraka (BSc,MSc Cand.)

Cloning in bacteria. Presenter: Vito Baraka (BSc,MSc Cand.) Cloning in bacteria Presenter: Vito Baraka (BSc,MSc Cand.) Introduction DNA cloning involves separating a specific gene or DNA segment from a larger chromosome, attaching it to a small molecule of carrier

More information

HiPer Transformation Teaching Kit

HiPer Transformation Teaching Kit HiPer Transformation Teaching Kit Product Code: HTBM017 Number of experiments that can be performed: 10 Duration of Experiment: 4 days Day 1- Preparation of media and revival of E. coli Host Day 2- Inoculation

More information

AP Biology Gene Expression/Biotechnology REVIEW

AP 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 information

GeNei TM Transformation Teaching Kit Manual

GeNei TM Transformation Teaching Kit Manual Teaching Kit Manual Cat No. New Cat No. KT07 107385 KT07A 106220 Revision No.: 00060505 CONTENTS Page No. Objective 3 Principle 3 Kit Description 6 Materials Provided 7 Procedure 9 Observation & Interpretation

More information

Genetics Lecture Notes Lectures 17 19

Genetics Lecture Notes Lectures 17 19 Genetics Lecture Notes 7.03 2005 Lectures 17 19 Lecture 17 Gene Regulation We are now going to look at ways that genetics can be used to study gene regulation. The issue is how cells adjust the expression

More information

Gene Cloning & DNA Analysis

Gene Cloning & DNA Analysis CSS451 CSS/HRT 451 Gene Cloning & DNA Analysis Chapter 4-5 T-DNA LB auxin cytokin opine Oncogenic genes RB vir genes ori opine catabolism Guo-qing Song Part 1 Basic principles Gene Cloning & DNA Analysis

More information

Molecular Cell Biology - Problem Drill 11: Recombinant DNA

Molecular 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 information

Chapter 9 Genetic Engineering

Chapter 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 information

Lecture 25 (11/15/17)

Lecture 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 information

GENE REGULATION IN PROKARYOTES

GENE REGULATION IN PROKARYOTES GENE REGULATION IN PROKARYOTES Prepared by Brenda Leady, University of Toledo Copyright (c) The McGraw-Hill Companies, Inc. Permission required for reproduction or display. 1 Gene regulation refers to

More information

DNA Structure and Properties Basic Properties Predicting Melting Temperature. Dinesh Yadav

DNA Structure and Properties Basic Properties Predicting Melting Temperature. Dinesh Yadav DNA Structure and Properties Basic Properties Predicting Melting Temperature Dinesh Yadav Nucleic Acid Structure Question: Is this RNA or DNA? Molecules of Life, pp. 15 2 Nucleic Acid Bases Molecules of

More information

Chapter 9. Biotechnology and DNA Technology

Chapter 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 information

Name Per AP: CHAPTER 27: PROKARYOTES (Bacteria) p559,

Name Per AP: CHAPTER 27: PROKARYOTES (Bacteria) p559, AP: CHAPTER 27: PROKARYOTES (Bacteria) p559, 561-564 1. How does the bacterial chromosome compare to a eukaryotic chromosome? 2. What is a plasmid? 3. How fast can bacteria reproduce? 4. What is a bacterial

More information

7.1 The lac Operon 7-1

7.1 The lac Operon 7-1 7.1 The lac Operon The lac operon was the first operon discovered It contains 3 genes coding for E. coli proteins that permit the bacteria to use the sugar lactose Galactoside permease (lacy) which transports

More information

CHAPTER 2A HOW DO YOU BEGIN TO CLONE A GENE? CHAPTER 2A STUDENT GUIDE 2013 Amgen Foundation. All rights reserved.

CHAPTER 2A HOW DO YOU BEGIN TO CLONE A GENE? CHAPTER 2A STUDENT GUIDE 2013 Amgen Foundation. All rights reserved. CHAPTER 2A HOW DO YOU BEGIN TO CLONE A GENE? 35 INTRODUCTION In the Program Introduction, you learned that the increase in diabetes in the United States has resulted in a great demand for its treatment,

More information

The plasmid shown to the right has an oriv and orit at the positions indicated, and is known to replicate bidirectionally.

The plasmid shown to the right has an oriv and orit at the positions indicated, and is known to replicate bidirectionally. Name Microbial Genetics, BIO 410/510 2008 Exam II The plasmid shown to the right has an oriv and orit at the positions indicated, and is known to replicate bidirectionally. 1.) Indicate where replication

More information

Lectures of Dr.Mohammad Alfaham. The Bacterial Genetics

Lectures of Dr.Mohammad Alfaham. The Bacterial Genetics Lectures of Dr.Mohammad Alfaham The Bacterial Genetics is the total collection of genes carried by a bacterium both on its chromosome and on its extrachromosomal genetic elements (plasmids) A Gene A gene

More information

pbluescript II RI Predigested Vector

pbluescript II RI Predigested Vector pbluescript II RI Predigested Vector INSTRUCTION MANUAL Catalog #212250 Revision A For In Vitro Use Only 212250-12 LIMITED PRODUCT WARRANTY This warranty limits our liability to replacement of this product.

More information

Genetics and Genomics in Medicine Chapter 3. Questions & Answers

Genetics 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

BCH 462 Competent Cells Formation and Transformation of Competent Cells with plasmid DNA.

BCH 462 Competent Cells Formation and Transformation of Competent Cells with plasmid DNA. Lab#2 BCH 462 Competent Cells Formation and Transformation of Competent Cells with plasmid DNA. Outlines: 1-Insertion of foreign gene to the plasmid. 2-Competent cell. 3-Transformation of bacterial cell.

More information

The GeneEditor TM in vitro Mutagenesis System: Site- Directed Mutagenesis Using Altered Beta-Lactamase Specificity

The GeneEditor TM in vitro Mutagenesis System: Site- Directed Mutagenesis Using Altered Beta-Lactamase Specificity Promega Notes Magazine Number 62, 1997, p. 02 The GeneEditor TM in vitro Mutagenesis System: Site- Directed Mutagenesis Using Altered Beta-Lactamase Specificity By Christine Andrews and Scott Lesley Promega

More information

Introducing new DNA into the genome requires cloning the donor sequence, delivery of the cloned DNA into the cell, and integration into the genome.

Introducing new DNA into the genome requires cloning the donor sequence, delivery of the cloned DNA into the cell, and integration into the genome. Key Terms Chapter 32: Genetic Engineering Cloning describes propagation of a DNA sequence by incorporating it into a hybrid construct that can be replicated in a host cell. A cloning vector is a plasmid

More information

Biotechnology and Energy Conservation. Prof. Dr.oec.troph. Ir. Krishna Purnawan Candra, M.S. Program Magister Ilmu Lingkungan Universitas Mulawarman

Biotechnology and Energy Conservation. Prof. Dr.oec.troph. Ir. Krishna Purnawan Candra, M.S. Program Magister Ilmu Lingkungan Universitas Mulawarman Biotechnology and Energy Conservation Prof. Dr.oec.troph. Ir. Krishna Purnawan Candra, M.S. Program Magister Ilmu Lingkungan Universitas Mulawarman 12 th Lecture Genetic Engineering The Aim: Students can

More information

Cloning Vectors Ameer Effat M. Elfarash

Cloning Vectors Ameer Effat M. Elfarash Cloning Vectors Ameer Effat M. Elfarash Dept. of Genetics Fac. of Agriculture, Assiut Univ. amir_effat@yahoo.com CLONING VECTORS Cloning vectors are DNA molecules that are used to "transport" cloned sequences

More information

The Regulation of Bacterial Gene Expression

The Regulation of Bacterial Gene Expression The Regulation of Bacterial Gene Expression Constitutive genes are expressed at a fixed rate Other genes are expressed only as needed Inducible genes Repressible genes Catabolite repression Pre-transcriptional

More information

Microbiology 微生物学 Spring-Summer

Microbiology 微生物学 Spring-Summer Microbiology 微生物学 2017 Spring-Summer Relevant Information and Resources Course slides can be found at http://mypage.zju.edu.cn/haichun 教学工作 Course-related questions will be answered through emails. Textbook:

More information

number Done by Corrected by Doctor Hamed Al Zoubi

number Done by Corrected by Doctor Hamed Al Zoubi number 3 Done by Neda a Baniata Corrected by Waseem Abu Obeida Doctor Hamed Al Zoubi Note: it is important to refer to slides. Bacterial genetics *The main concepts we will talk about in this lecture:

More information

CHEM 4420 Exam I Spring 2013 Page 1 of 6

CHEM 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 information

MOLECULAR GENETICS: TRANSFORMATION AND CLONING adapted by Dr. D. L. Vogelien

MOLECULAR GENETICS: TRANSFORMATION AND CLONING adapted by Dr. D. L. Vogelien Introduction MOLECULAR GENETICS: TRANSFORMATION AND CLONING adapted by Dr. D. L. Vogelien The field of molecular genetics has resulted in a number of practical applications that have been of tremendous

More information

Lecture 9 Controlling gene expression

Lecture 9 Controlling gene expression Lecture 9 Controlling gene expression BIOLOGY Campbell, Reece and Mitchell Chapter 18 334- (352-356) Every cell in your body contains the same number of genes approximately 35, 000 DNA is wound around

More information

REGULATION OF GENE EXPRESSION

REGULATION OF GENE EXPRESSION REGULATION OF GENE EXPRESSION Each cell of a living organism contains thousands of genes. But all genes do not function at a time. Genes function according to requirements of the cell. Genes control the

More information

Lecture 3 (FW) January 28, 2009 Cloning of DNA; PCR amplification Reading assignment: Cloning, ; ; 330 PCR, ; 329.

Lecture 3 (FW) January 28, 2009 Cloning of DNA; PCR amplification Reading assignment: Cloning, ; ; 330 PCR, ; 329. Lecture 3 (FW) January 28, 2009 Cloning of DNA; PCR amplification Reading assignment: Cloning, 240-245; 286-87; 330 PCR, 270-274; 329. Take Home Lesson(s) from Lecture 2: 1. DNA is a double helix of complementary

More information

Biology 201 (Genetics) Exam #3 120 points 20 November Read the question carefully before answering. Think before you write.

Biology 201 (Genetics) Exam #3 120 points 20 November Read the question carefully before answering. Think before you write. Name KEY Section Biology 201 (Genetics) Exam #3 120 points 20 November 2006 Read the question carefully before answering. Think before you write. You will have up to 50 minutes to take this exam. After

More information

Genetic Engineering & Recombinant DNA

Genetic 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 information

M I C R O B I O L O G Y WITH DISEASES BY TAXONOMY, THIRD EDITION

M I C R O B I O L O G Y WITH DISEASES BY TAXONOMY, THIRD EDITION M I C R O B I O L O G Y WITH DISEASES BY TAXONOMY, THIRD EDITION Chapter 7 Microbial Genetics Lecture prepared by Mindy Miller-Kittrell, University of Tennessee, Knoxville The Structure and Replication

More information

Problem Set 8. Answer Key

Problem Set 8. Answer Key MCB 102 University of California, Berkeley August 11, 2009 Isabelle Philipp Online Document Problem Set 8 Answer Key 1. The Genetic Code (a) Are all amino acids encoded by the same number of codons? no

More information

GENE CLONING: overview

GENE CLONING: overview TMMO504: Laboratory Molecular Tropical Medicine and Genetics GENE CLONING: overview Instructor: Asst.Prof.Dr. Santi Maneewatchararangsri Department of Molecular Tropical Medicine and Genetics, Mahidol

More information

Biotechnology. Chapter 20. Biology Eighth Edition Neil Campbell and Jane Reece. PowerPoint Lecture Presentations for

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 information

3 Designing Primers for Site-Directed Mutagenesis

3 Designing Primers for Site-Directed Mutagenesis 3 Designing Primers for Site-Directed Mutagenesis 3.1 Learning Objectives During the next two labs you will learn the basics of site-directed mutagenesis: you will design primers for the mutants you designed

More information

CHAPTER 13 LECTURE SLIDES

CHAPTER 13 LECTURE SLIDES CHAPTER 13 LECTURE SLIDES Prepared by Brenda Leady University of Toledo To run the animations you must be in Slideshow View. Use the buttons on the animation to play, pause, and turn audio/text on or off.

More information

Vectors for Gene Cloning: Plasmids and Bacteriophages

Vectors for Gene Cloning: Plasmids and Bacteriophages Vectors for Gene Cloning: Plasmids and Bacteriophages DNA molecule must be able to replicate within the host cell to be able to act as a vector for gene cloning, so that numerous copies of the recombinant

More information

Heterologous protein expression systems

Heterologous protein expression systems IMBB-FORTH Heterologous protein expression systems What are they? Protein expression systems producing desired polypeptides from recombinant genes. Plasmids carry and express the desired recombinant genes

More information

Genetics Lecture 21 Recombinant DNA

Genetics 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 information

AP Biology. Chapter 20. Biotechnology: DNA Technology & Genomics. Biotechnology. The BIG Questions. Evolution & breeding of food plants

AP 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 information

Regulation of gene expression. (Lehninger pg )

Regulation of gene expression. (Lehninger pg ) Regulation of gene expression (Lehninger pg. 1072-1085) Today s lecture Gene expression Constitutive, inducible, repressible genes Specificity factors, activators, repressors Negative and positive gene

More information

Chapter 4: How Cells Work

Chapter 4: How Cells Work Chapter 4: How Cells Work David Shonnard Department of Chemical Engineering 1 Presentation Outline: l l l l l Introduction : Central Dogma DNA Replication: Preserving and Propagating DNA Transcription:

More information

Recombinant DNA Technology. The Role of Recombinant DNA Technology in Biotechnology. yeast. Biotechnology. Recombinant DNA technology.

Recombinant 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 information

Green Fluorescent Protein (GFP) Purification. Hydrophobic Interaction Chromatography

Green Fluorescent Protein (GFP) Purification. Hydrophobic Interaction Chromatography Green Fluorescent Protein (GFP) Purification Hydrophobic Interaction Chromatography What is the GFP gene? GFP is a green fluorescent protein that is normally found in jellyfish. It has been engineered

More information

BIOLOGY LTF DIAGNOSTIC TEST DNA to PROTEIN & BIOTECHNOLOGY

BIOLOGY LTF DIAGNOSTIC TEST DNA to PROTEIN & BIOTECHNOLOGY Biology Multiple Choice 016074 BIOLOGY LTF DIAGNOSTIC TEST DNA to PROTEIN & BIOTECHNOLOGY Test Code: 016074 Directions: Each of the questions or incomplete statements below is followed by five suggested

More information

Certificate of Analysis

Certificate of Analysis Certificate of Analysis pet6xhn Expression Vector Set Contents Product Information... 1 pet6xhn-n, pet6xhn-c, and pet6xhn-gfpuv Vector Information... 2 Location of Features... 4 Additional Information...

More information

1. What is the structure and function of DNA? Describe in words or a drawing the structure of a DNA molecule. Be as detailed as possible.

1. What is the structure and function of DNA? Describe in words or a drawing the structure of a DNA molecule. Be as detailed as possible. INTRODUCTION In the Program Introduction, you learned that the increase in diabetes in the United States has resulted in a great demand for its treatment, insulin. You also learned that the best way to

More information

Computational Biology I LSM5191

Computational 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 information

EcoR1 is a type IIP restriction endonuclease which cleaves the palindromic

EcoR1 is a type IIP restriction endonuclease which cleaves the palindromic Transfer of the Fungal cdna CIH-1 from the Plasmid Vector pbk CMV to the Plasmid Vector puc19 and sub- Cloning Mediated Recombinant puc19 Amplification INTRODUCTION Molecular cloning is a method used for

More information

Biology (Microbiology): Exam #3

Biology (Microbiology): Exam #3 NAME: PLEDGE: Biology 50-384 (Microbiology): Exam #3 1. You have isolated a series of mutants that have altered patterns of ß-galactosidase and lactose permease activity (i.e. they are different that the

More information

DNA Structure and Properties

DNA Structure and Properties DNA Structure and Properties Biochemistry Boot Camp 2017 Session #6 Nick Fitzkee nfitzkee@chemistry.msstate.edu DNA DNA- a polymer of deoxyribonucleotides Found in chromosomes, mitochondria and chloroplasts

More information

How Do You Clone a Gene?

How Do You Clone a Gene? S-20 Edvo-Kit #S-20 How Do You Clone a Gene? Experiment Objective: The objective of this experiment is to gain an understanding of the structure of DNA, a genetically engineered clone, and how genes are

More information

Sequence Analysis Lab Protocol

Sequence Analysis Lab Protocol Sequence Analysis Lab Protocol You will need this handout of instructions The sequence of your plasmid from the ABI The Accession number for Lambda DNA J02459 The Accession number for puc 18 is L09136

More information

Name_BS50 Exam 3 Key (Fall 2005) Page 2 of 5

Name_BS50 Exam 3 Key (Fall 2005) Page 2 of 5 Name_BS50 Exam 3 Key (Fall 2005) Page 2 of 5 Question 1. (14 points) Several Hfr strains derived from the same F + strain were crossed separately to an F - strain, giving the results indicated in the table

More information

HI-Control BL21(DE3) & HI-Control 10G Chemically Competent Cells

HI-Control BL21(DE3) & HI-Control 10G Chemically Competent Cells HI-Control BL21(DE3) & HI-Control 10G Chemically Competent Cells FOR RESEARCH USE ONLY. NOT FOR HUMAN OR DIAGNOSTIC USE MA156 Rev.31OCT2016 Table of Contents Components & Storage Conditions... 3 HI-Control

More information

1. Page 90: Cellular Metabolism Explain what the everyday use of the word metabolism means to you.

1. Page 90: Cellular Metabolism Explain what the everyday use of the word metabolism means to you. Biology 100 Winter 2013 North Seattle Community College Reading Guide 10 Metabolism, Enzymes, and Building a Protein Reading: 1) Chapter 5 (various pages) in Microbiology Demystified 2) Chapter 7 (various

More information

Transforming Bacteria

Transforming Bacteria Properties of E. coli Strains for Subcloning Common laboratory strains of E. coli, like JM109, DH5α, and XL-1 Blue, are different from their wildtype counterparts. These strains carry some mutations designed

More information

Name 10 Molecular Biology of the Gene Test Date Study Guide You must know: The structure of DNA. The major steps to replication.

Name 10 Molecular Biology of the Gene Test Date Study Guide You must know: The structure of DNA. The major steps to replication. Name 10 Molecular Biology of the Gene Test Date Study Guide You must know: The structure of DNA. The major steps to replication. The difference between replication, transcription, and translation. How

More information

I. Prokaryotic Gene Regulation. Figure 1: Operon. Operon:

I. Prokaryotic Gene Regulation. Figure 1: Operon. Operon: I. Prokaryotic Gene Regulation Figure 1: Operon Operon: a) Regulatory Elements consist of an Operator that serves as the on-off switch for the genes of the operon. Also contains a promoter for the Structural

More information

7 Gene Isolation and Analysis of Multiple

7 Gene Isolation and Analysis of Multiple Genetic Techniques for Biological Research Corinne A. Michels Copyright q 2002 John Wiley & Sons, Ltd ISBNs: 0-471-89921-6 (Hardback); 0-470-84662-3 (Electronic) 7 Gene Isolation and Analysis of Multiple

More information

BA, BSc, and MSc Degree Examinations

BA, BSc, and MSc Degree Examinations Examination Candidate Number: Desk Number: BA, BSc, and MSc Degree Examinations 2017-8 Department : BIOLOGY Title of Exam: Genetics Time Allowed: 1 hour and 30 minutes Marking Scheme: Total marks available

More information

A Discovery Laboratory Investigating Bacterial Gene Regulation

A Discovery Laboratory Investigating Bacterial Gene Regulation Chapter 8 A Discovery Laboratory Investigating Bacterial Gene Regulation Robert Moss Wofford College 429 N. Church Street Spartanburg, SC 29307 mosssre@wofford.edu Bob Moss is an Associate Professor of

More information

Sample Question Paper Session Class XII Biotechnology (045) Marking Scheme

Sample Question Paper Session Class XII Biotechnology (045) Marking Scheme Sample Question Paper Session 2015-16 Class XII Biotechnology (045) Marking Scheme SECTION-A 1. Methylation Student will add a methyl group to one or two bases within the sequence recognized by enzyme.

More information

Chapter 18: Regulation of Gene Expression. 1. Gene Regulation in Bacteria 2. Gene Regulation in Eukaryotes 3. Gene Regulation & Cancer

Chapter 18: Regulation of Gene Expression. 1. Gene Regulation in Bacteria 2. Gene Regulation in Eukaryotes 3. Gene Regulation & Cancer Chapter 18: Regulation of Gene Expression 1. Gene Regulation in Bacteria 2. Gene Regulation in Eukaryotes 3. Gene Regulation & Cancer Gene Regulation Gene regulation refers to all aspects of controlling

More information

Justin Veazey. Experiment 3; Analysis of digestion products of puc19, GFPuv, and pgem-t easy

Justin Veazey. Experiment 3; Analysis of digestion products of puc19, GFPuv, and pgem-t easy Veazey 1 Justin Veazey 7A Experiment 3; Analysis of digestion products of puc19, GFPuv, and pgem-t easy Construction of recombinants GFPuv-pGEM-T easy and GFPuv-pUC19 Transformation and analysis of recombinant

More information

LAB #14: Rapid Colony Transformation of E. coli with Plasmid DNA

LAB #14: Rapid Colony Transformation of E. coli with Plasmid DNA LAB #14: Rapid Colony Transformation of E. coli with Plasmid DNA Objective: In this laboratory investigation, plasmids containing fragments of foreign DNA will be used to transform Escherichia coli cells,

More information

BIOLOGY. Chapter 16 GenesExpression

BIOLOGY. Chapter 16 GenesExpression BIOLOGY Chapter 16 GenesExpression CAMPBELL BIOLOGY TENTH EDITION Reece Urry Cain Wasserman Minorsky Jackson 18 Gene Expression 2014 Pearson Education, Inc. Figure 16.1 Differential Gene Expression results

More information

Site-directed mutagenesis of proteins

Site-directed mutagenesis of proteins IFM/Kemi Linköpings Universitet August 2013/LGM Labmanual Site-directed mutagenesis of proteins Figur 1: Flow-chart of the site-directed mutagenesis lab exercise 2 Site-specific mutagenesis Introduction

More information

encodes a sigma factor to modify the recognition of the E.coli RNA polymerase (Several other answers would also be acceptable for each phage)

encodes a sigma factor to modify the recognition of the E.coli RNA polymerase (Several other answers would also be acceptable for each phage) Name Student ID# Bacterial Genetics, BIO 4443/6443 Spring Semester 2001 Final Exam 1.) Different bacteriophage utilize different mechanisms to ensure that their own genes (and not host s genes) are transcribed

More information

2054, Chap. 13, page 1

2054, Chap. 13, page 1 2054, Chap. 13, page 1 I. Microbial Recombination and Plasmids (Chapter 13) A. recombination = process of combining genetic material from 2 organisms to produce a genotype different from either parent

More information

DNA Technology. Asilomar Singer, Zinder, Brenner, Berg

DNA 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

Some types of Mutagenesis

Some types of Mutagenesis Mutagenesis What Is a Mutation? Genetic information is encoded by the sequence of the nucleotide bases in DNA of the gene. The four nucleotides are: adenine (A), thymine (T), guanine (G), and cytosine

More information

Student Manual. pglo Transformation

Student Manual. pglo Transformation Student Manual pglo Transformation Lesson 1 Introduction to Transformation In this lab you will perform a procedure known as genetic transformation. Remember that a gene is a piece of DNA which provides

More information

MCB 421 First Exam October 4, 2004

MCB 421 First Exam October 4, 2004 1. (10 pts). An E. coli strain (strain A) that lacks an inducing prophage and carries the F factor is heavily irradiated with UV light and then mixed 1:1 with a second E. coli strain (strain B) that carries

More information

Relevance of Primer Design to the Effective Disruption of the laci gene in Escherichia coli C29 using the Lambda Red Recombinase System

Relevance of Primer Design to the Effective Disruption of the laci gene in Escherichia coli C29 using the Lambda Red Recombinase System Relevance of Primer Design to the Effective Disruption of the laci gene in Escherichia coli C29 using the Lambda Red Recombinase System JADA BROEKHUIZEN, OMEED HADISFAR, DAVID LIU, JAMES MCFARLANE, AND

More information

ONTARIO SCIENCE CENTRE. Teacher Guide. Way to Glow Program

ONTARIO SCIENCE CENTRE. Teacher Guide. Way to Glow Program ONTARIO SCIENCE CENTRE Teacher Guide Way to Glow Program Table of Contents Bacterial transformation background information 3 Experimental procedure 5 Expected results 7 Post-program activity sheet 8 Post-program

More information

Regulation of Gene Expression

Regulation of Gene Expression Slide 1 Chapter 18 Regulation of Gene Expression PowerPoint Lecture Presentations for Biology Eighth Edition Neil Campbell and Jane Reece Lectures by Chris Romero, updated by Erin Barley with contributions

More information

BIOTECHNOLOGY OLD BIOTECHNOLOGY (TRADITIONAL BIOTECHNOLOGY) MODERN BIOTECHNOLOGY RECOMBINANT DNA TECHNOLOGY.

BIOTECHNOLOGY OLD BIOTECHNOLOGY (TRADITIONAL BIOTECHNOLOGY) MODERN BIOTECHNOLOGY RECOMBINANT DNA TECHNOLOGY. BIOTECHNOLOGY Biotechnology can be defined as the use of micro-organisms, plant or animal cells or their components or enzymes from organisms to produce products and processes (services) useful to human

More information

3. human genomics clone genes associated with genetic disorders. 4. many projects generate ordered clones that cover genome

3. human genomics clone genes associated with genetic disorders. 4. many projects generate ordered clones that cover genome Lectures 30 and 31 Genome analysis I. Genome analysis A. two general areas 1. structural 2. functional B. genome projects a status report 1. 1 st sequenced: several viral genomes 2. mitochondria and chloroplasts

More information

Section A: Prokaryotes Types and Structure 1. What is microbiology?

Section A: Prokaryotes Types and Structure 1. What is microbiology? Section A: Prokaryotes Types and Structure 1. What is microbiology? 2. Compare and contrast characteristics of each bacterial type: Eubacteria and Archaebacteria. Eubacteria Both Archaebacteria 3. Label

More information

Independent Study Guide The Blueprint of Life, from DNA to Protein (Chapter 7)

Independent Study Guide The Blueprint of Life, from DNA to Protein (Chapter 7) Independent Study Guide The Blueprint of Life, from DNA to Protein (Chapter 7) I. General Principles (Chapter 7 introduction) a. Morse code distinct series of dots and dashes encode the 26 letters of the

More information

Read the question carefully before answering. Think before you write. If I can not read your handwriting, I will count the question wrong.

Read the question carefully before answering. Think before you write. If I can not read your handwriting, I will count the question wrong. Name KEY Note Total points added up to only 98 points so everyone received 2 free points to make total points 100. Biology 201 (Genetics) Exam #3 23 November 2004 Read the question carefully before answering.

More information

BC2004 Review Sheet for Lab Exercises 7-11 Spring Semester 2005

BC2004 Review Sheet for Lab Exercises 7-11 Spring Semester 2005 BC2004 Review Sheet for Lab Exercises 7-11 Spring Semester 2005 Lab Exercise 7 Drosophila crosses, three weeks Vocabulary: phenotype, genotype, gene, allele, locus (loci), sex chromosomes, autosomes, homozygous,

More information

Supporting Information-Tables

Supporting Information-Tables Supporting Information-Tables Table S1. Bacterial strains and plasmids used in this work Bacterial strains Description Source of reference Streptococcus pneumoniae 1 Cp1015 non-capsulated and βl susceptible

More information

Chapter 20: Biotechnology

Chapter 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 information

Molecular Techniques Third-year Biology

Molecular Techniques Third-year Biology PLANNING Genetics Lab practices Molecular Techniques. Genetics Lab practices protocol. 2015-16 PCR-DIRECTED MUTAGENESIS, MOLECULAR CLONING AND RESTRICTION ANALYSIS Sessions 1 & 2 (2x3 hours): PCR-directed

More information

Biology Notes (Term 3!)

Biology Notes (Term 3!) Biology Notes (Term 3!) --------------------------------------------------------------------------------------------------------------------------- RNA Definition Ribonucleic acid (RNA) is another nucleic

More information

GENE REGULATION slide shows by Kim Foglia modified Slides with blue edges are Kim s

GENE REGULATION slide shows by Kim Foglia modified Slides with blue edges are Kim s GENE REGULATION slide shows by Kim Foglia modified Slides with blue edges are Kim s 2007-2008 Bacterial metabolism Bacteria need to respond quickly to changes in their environment STOP GO if they have

More information