Confirming the Phenotypes of E. coli Strains
|
|
- Emma Hodges
- 6 years ago
- Views:
Transcription
1 Confirming the Phenotypes of E. coli Strains INTRODUCTION Before undertaking any experiments, we need to confirm that the phenotypes of the E. coli strains we intend to use in the planned experiments correspond with their published genotypes. This is an essential preamble to any bacterial genetics project. More often than not, phenotypes in bacterial genetics are evaluated by plating cells on agar plates containing specially formulated growth media. Note that here we are using the techniques of classical genetics do not consult genotype directly, but only indirectly, via phenotype. Directly verifying the genotype involves Molecular Biology rather than Genetics. This means using techniques such as cloning, DNA sequencing, PCR, or Southern analysis. While these methods potentially provide a more definitive characterization of a strains genotype, they all generally involve more time and money than straightforward phenotypic characterization. So, it's a trade-off. STRAIN DESCRIPTION Here is a list of the 4 strains we will check, and a description of their genotypes and properties. Following that there will be a description of the growth media. We want to help guide you through the process of putting these two sets of descriptions together in your heads so that you understand what each strain should do on each of the media. UCSC Stain# SC071 SC142 This is MG1655, the standard "wild-type" for the E. coli K-12 lineage. It is F - (λ - ), but otherwise should have the "wild-type" genotype for all genes of interest to us in the course. Δ(gpt-lac) SC109 Δ(gpt-lac) rpsl (=stra1) SC138 Δ(gpt-lac) Tn5 F' proa, prob laci, lacz, lacy Page 1 of 6
2 Δ(gpt-lac) was described in the nomenclature exercise. The rpsl in strain 109 is the classic mutant allele originally designated stra1; this was also described in the nomenclature exercise. SC138 is a more complicated beast. For starters, it has a copy of the transposon Tn5 inserted in the chromosome at an unknown location. You will need to read the posted references on Tn5 to find out how it will affect the behavior of SC138. SC138 also has an F' plasmid. F' plasmids are derivatives of the original F conjugal plasmid that has acquired one ore more genes from the E. coli genome. In this F', the plasmid has carries a large region of the E. coli genome that extends from the genes proa and prob ( ) to the Lac Operon ( ). Description of Media Broth Media vs. Minimal Media Bacterial growth media fall into 2 broad categories, BROTH Media and MINIMAL Media. It is critical to understand the distinction between these types in order to interpret your results. BROTH media are generally prepared from aqueous extracts of cells or tissues, combined with hydrolyzed proteins. Thus, they provide a rich variety of organic molecules (amino acids, nucleotides, carbohydrates, etc.) that can be used as carbon and energy sources for bacterial growth. They support rapid growth, even of E. coli strains with mutations that eliminate some of the metabolic characteristics of the wild-type. They are also easy to prepare. The drawback is that their molecular composition is not only complex, but undefined. the broth medium we use is medium, the ubiquitous, standard E. coli growth medium in molecular biology. MINIMAL media may contain only a single type of organic molecule added to a solution of inorganic salts (ammonium, phosphate, sulfate, etc.). Growth of a strain in a MINIMAL medium implies that the strain is prototrophic, meaning that it has a complete set of biosynthetic pathways, and that it can metabolize the particular carbon source provided (frequently glucose or some other sugar). Auxotrophic mutants cannot grow in un-supplemented MINIMAL media because they lack at least one complete biosynthetic pathway. For example, an E. coli mutant strain lacking a gene for an enzyme in the histidine biosynthetic pathway has a His - phenotype and will not grow in Glucose Minimal Medium unless the medium is supplemented with histidine. Minimal media are more trouble to prepare, and do not support as rapid growth as broth media, but they have the great advantage that their molecular composition is defined. A defined medium is one whose composition is known precisely at the molecular level. Defined media are almost always used in metabolic, nutritional or physiological growth experiments. The media used, and the plate code used to identify them are: Broth Media: + Streptomycin I + Rifampicin II + Kanamycin K + Triclosan Ts + X-Gal X* Minimal Media: Minimal Glucose I I Minimal Glucose + Proline I I P Minimal Lactose + Proline I II P Page 2 of 6
3 *X-Gal, an artificial, chromogenic substrate for the enzyme beta-galactosidase (the lacz gene product). X-Gal is colorless, but its hydrolysis product is blue. Therefore, only Lac + strains produce blue colonies on X-Gal medium. This is the substrate commonly used in detecting recombinant plasmids in classic molecular cloning. You should print a copy of the Agar Plate Code from the web site and keep it in your notebook. Page 3 of 6
4 PROCEDURE DAY 1 We provide you with overnight cultures of the 4 strains. Overnight cultures will be a typical starting point for many of our exercises. During the growth of these cultures (15-24 hrs) one or more essential nutrients become/s depleted, and the growth rate falls to an insignificant level. That is, the cell density in an overnight culture probably represents the carrying capacity of the medium, or, as a microbiologist would say, the culture is in stationary phase. For E. coli cultures grown in Medium, the cell concentration in an overnight culture is usually about 2 X 10 9 cells/ml. Compare the appearance of your overnight cultures to the appearance of sterile Medium. After your overnight cultures sit undisturbed in the bench for an hour or so look at them again. How does the appearance of the culture change? What does this mean? Dilute 10 ul of each overnight culture into 1 ml of sterile saline (0.9% NaCl) and mix thoroughly. (In 4 separate tubes, that is. Don't mix all the different strains together in one tube.) This represents a 10-2 dilution. 0.9% NaCl (sterile saline) is a generic vehicle for the suspension and dilution of E. coli. No nutrients are provided for growth, but isotonic conditions prevent the osmotic effects that might injure the cells if they were diluted in di water. Get all your agar plates and label them on the bottom according to the diagram below. Transfer 5 ul drops of each diluted overnight culture onto the agar plates. (Do one strain on all plates using the same micropipette tip. Then change tips and do the next strain.) Streak the droplets lengthwise on the plates with a sterile glass rod. See the diagram below. (You can streak the same strain on all the media before you need to re-sterilize it.) Allow the plates to sit on the bench until all free liquid has been absorbed into the agar. Incubate the agar plates upside down in a 37 C incubator. Wild-type E. coli produce visible colonies (>0.5 mm dia.) on plates overnight. Formation of visible colonies on minimal plates requires incubation for hours. Page 4 of 6
5 PROCEDURE DAY 2 Before you come to class, complete the table below by filling in your predicted result for the 5 strains on each medium. Describe your prediction in terms of what you would see when you look at the plate. Be sure to define any abbreviations or symbols (i.e. + = confluent growth etc.). Make a copy of the completed table and give one to the TA before you examine your plates. Keep the other copy for reference. Don't fret about "getting it wrong", just take your best shot and, certainly, ask us questions that will help you figure out what should be happening. Strain Strep Kan Rif Tric X-Gal Minimal Glucose Minimal Glucose Proline Minimal Lactose Proline SC071 SC142 SC109 SC138 Now, examine your plates and carefully record your observations in your notebook. Words and pictures can both contribute to robust results. Page 5 of 6
6 ASSIGNMENT 1. [10 pts.] Completed table with your predicted results (submitted before making your observations.) 2. [20 pts.] Completed the table with the actual, observed result for the 4 strains on each medium. Note: The results may not be as simple as "Growth" vs No Growth". 2. [40 pts.] Write an organized discussion of whether or not the phenotypes of the strains, as demonstrated by growth on the agar plates, are consistent with their reputed genotypes. In particular, focus on an analysis of any discrepancies between your results and your original predictions. This is an opportunity to use the strategy of first creating an outline of what you wish to convey, so that you can avoid a "stream of consciousness" effect. 3. [10 pts.] Approximately how many E.coli cells were in each 10 ul drop spotted on the plates? 4. [10 pts.] Why did we dilute the overnight cultures in sterile saline before plating them? Wouldn't it have been faster to just plate them directly? Page 6 of 6
Biology 322 Fall 2010 Transfer of genetic information in the bacterium Escherichia coli: Part I
Biology 322 Fall 2010 Transfer of genetic information in the bacterium Escherichia coli: Part I REQUIRED Reading Assignments: Superbugs on the Hoof http://fire.biol.wwu.edu/trent/trent/superbugs.pdf Triple
More information7.02 Microbial Genetics in Lab Quiz. Fall, September 27, 2001 ANSWER KEY
7.02 Microbial Genetics in Lab Quiz Fall, 2001 September 27, 2001 ANSWER KEY This quiz contains 4 questions worth a total of 48 points. Be sure to write your name, Bench letter and Undergraduate TA s (UTA)
More informationHOUR EXAM I BIOLOGY 422 FALL, In the spirit of the honor code, I pledge that I have neither given nor received help on this exam.
Name First Last (Please Print) PID Number - HOUR EXAM I BIOLOGY 422 FALL, 2011 In the spirit of the honor code, I pledge that I have neither given nor received help on this exam. 1 Signature 2 3 4 5 6
More informationFigure 1. Map of cloning vector pgem T-Easy (bacterial plasmid DNA)
Texas A&M University-Corpus Christi CHEM4402 Biochemistry II Laboratory Laboratory 6: Ligation & Bacterial Transformation (Bring your text and laptop to class if you wish to work on your assignment during
More informationBACTERIAL CONJUGATION. To demonstrate the technical procedure to monitor the conjugational transfer of genetic material from one cell to another.
BACTERIAL CONJUGATION I. OBJECTIVES To demonstrate the technical procedure to monitor the conjugational transfer of genetic material from one cell to another. To learn about the various genetic elements
More informationHiPer 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 informationBIO440 Genetics Laboratory Transformation
BIO440 Genetics Laboratory Transformation The transfer of genetic information between bacteria has been occurring for billions of years. Humans first noticed this process in the laboratory in the 1920
More informationANSWERS TO Problem set questions from Exam 2 Unit Mutations, Bacterial Genetics, and Bacterial Gene Regulation
ANSWERS TO Problem set questions from Exam 2 Unit Mutations, Bacterial Genetics, and Bacterial Gene Regulation Central Dogma, Mutagens and Mutations 1. The three stop codons in the genetic code are 5 UAG3,
More informationSolutions to 7.02 Quiz III
Solutions to 7.02 Quiz III Class Average = 79 Standard Deviation = 12 Range Grade % 85-100 A 40 72-84 B 37 55-71 C 20 > 54 D/F 3 Question 1 On day 1 of the genetics lab, the entire 7.02 class did a transposon
More informationGenetics IV: Biochemical Genetics
Genetics IV: Biochemical Genetics 1. Population Genetics This field was advanced by laws proposed by two people Hardy and Weinberg (1908) Suppose in a population there are 2 alleles for a given gene, A
More informationThe 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 informationStep 1: Digest vector with Reason for Step 1. Step 2: Digest T4 genomic DNA with Reason for Step 2: Step 3: Reason for Step 3:
Biol/Chem 475 Spring 2007 Study Problems for Quiz 2 Quiz 2 (~50 pts) is scheduled for Monday May 14 It will cover all handouts and lab exercises to date except the handout/worksheet (yet to be distributed)
More informationA 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 informationFrom Kendall- idea that this lecture is like a microcosm of the whole course
411-2 2008 From Kendall- idea that this lecture is like a microcosm of the whole course Before starting: Nomenclature: Lac - phenotype vs IacZ or lacy genotype- phenotype is what's observed; genotype is
More informationBiology Lab Activity 4-5 DNA Transformation
Biology Lab Activity 4-5 DNA Transformation Scientists can insert genes into bacteria. The genes inserted in the Indo-Blu process (this lab) are on a circular piece of DNA called a plasmid. (The plasmid
More informationBACTERIAL GENETICS. How does the DNA in the bacterial cell replicate
BACTERIAL GENETICS Bacterial genetics is the study of gene structure and function in bacteria. Genetics itself is concerned with determining the number, location, and character of the genes of an organism.
More informationBiology 2250 Transformation laboratory
Biology 2250 Transformation laboratory Prior to lab: Discuss how to use micropipettes. Discuss how to use microcentrifuge balance. Discuss how to spread bacteria with alcohol and glass rods. Bacteria are
More informationBACTERIAL GENETICS: Labs I & II
BACTERIAL GENETICS: Labs I & II The Bacterial Genetics Labs will extend over two laboratory periods. During the first lab, you will set up two different experiments using the bacterium Escherichia coli.
More informationLABORATORY 5: TRANSFORMING BACTERIA WITH THE LIGATION PRODUCTS
LABORATORY 5: TRANSFORMING BACTERIA WITH THE LIGATION PRODUCTS So far in your quest to clone a gene you have produced recombinant plasmids and verified that you made the para-r plasmid containing the rfp
More informationMOLECULAR 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 informationAdvantages of genetic analysis in bacteria/phage. Selections vs Screens. Mutations. Mutations lecture: March 4, 2009
Mutations lecture: March 4, 2009 1. Genetic analysis of bacteria: the whys, hows, and whats 2. Luria-Delbrück and beyond: we still care! 3. Analysis of essential genes! RNA pol, merodiploids and amber
More informationExploring STEAM with Transformation
Exploring STEAM with Transformation Thomas Cynkar Edvotek www.edvotek.com Follow @Edvotek EDVOTEK Biotech The Biotechnology Education Company Celebrating 30 years of science education! EDVOTEK Educatio
More informationLac Operon contains three structural genes and is controlled by the lac repressor: (1) LacY protein transports lactose into the cell.
Regulation of gene expression a. Expression of most genes can be turned off and on, usually by controlling the initiation of transcription. b. Lactose degradation in E. coli (Negative Control) Lac Operon
More informationBiology 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 informationGENETICS - CLUTCH CH.5 GENETICS OF BACTERIA AND VIRUSES.
!! www.clutchprep.com CONCEPT: WORKING WITH MICROORGANISMS Bacteria are easy to with in a laboratory setting They are fast dividing, take up little space, and are easily grown in a lab - Plating is when
More informationGenetics Lecture Notes Lectures 13 16
Genetics Lecture Notes 7.03 2005 Lectures 13 16 Lecture 13 Transposable elements Transposons are usually from 10 3 to 10 4 base pairs in length, depending on the transposon type. The key property of transposons
More informationMCB 421 Exam #1 (A) Fall There are 9 questions and 1 supplement on last page. Answer all 9 questions. Be sure your name is on each page
MCB 421 Exam #1 (A) Fall 2006 There are 9 questions and 1 supplement on last page. Answer all 9 questions. Be sure your name is on each page 1). (8 points) A Luria-Delbruck fluctuation test was done to
More informationGeNei 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 informationpglo Transformation 1. Do the genetic transformation. 2. Determine the degree of success in your efforts to genetically alter an organism.
Introduction to Transformation pg Transformation In this lab you will perform a procedure known as genetic transformation. Remember that a gene is a piece of DNA which provides the instructions for making
More information5.) Name and describe one gene product in E.coli that is associated with performing each step in the recombination process. (6pts)
Student ID# Bacterial Genetics, BIO 4443/6443 Spring Semester 2001 Exam II 1.) What is the primary difference between conjugative plasmids and mobilizable plasmids? What genes are typically found on conjugative
More informationTransformation of Escherichia coli With a Chimeric Plasmid
Transformation of Escherichia coli With a Chimeric Plasmid Now that we have generated recombinant molecules, we must next amplify them by inserting them into an acceptable host so that they may be analyzed
More informationBiology 2180 Laboratory #7. Bacterial Growth and Transformation
Biology 2180 Laboratory #7 Name Bacterial Growth and Transformation Introduction: Most aspects of molecular biology require the use of basic microbiological methods. This section is included for those
More informationMicro 411/Genome Sci411 Exam Three
Name Question 1. 20pts Diagram the initiation of recombination by RecBCD starting with double stranded DNA. Make sure to label all enzymes, the location of the Chi site and where RecA is required. A. 5
More informationLab 5/5a Transformation of E. coli with a Recombinant Plasmid
Lab 5/5a Transformation of E. coli with a Recombinant Plasmid Lab 2 Pre Lab Readiness Familiarity and Proper use of micropipettes Remember the 1 st and 2 nd stops Aseptic Technique Antibiotic Resistance
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 informationTranscriptional Regulation
Transcriptional Regulation Gene expression responds to environmental conditions. Some regulatory proteins are present at only 5 10 copies, whereas under certain conditions, the expression of these proteins
More information4 Mutant Hunts - To Select or to Screen (Perhaps Even by Brute Force)
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) 4 Mutant Hunts - To Select or to Screen
More informationR1 12 kb R1 4 kb R1. R1 10 kb R1 2 kb R1 4 kb R1
Bcor101 Sample questions Midterm 3 1. The maps of the sites for restriction enzyme EcoR1 (R1) in the wild type and mutated cystic fibrosis genes are shown below: Wild Type R1 12 kb R1 4 kb R1 _ _ CF probe
More informationGenetics 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 informationTransformation: Theory. Day 2: Transformation Relevant Book Sections
Day 2: Transformation Relevant Book Sections We will follow the protocols provided in various industry-standard kits, instead of the protocols described in these chapters, but the chapters provide good
More informationAP Biology Lab 6 MOLECULAR BIOLOGY
AP Biology Laboratory Date: Name and Period: AP Biology Lab 6 MOLECULAR BIOLOGY OVERVIEW In this lab you will investigate some basic principles of molecular biology: 1. Plasmids containing specific fragments
More informationModule Code: BIO00007C
Examination Candidate Number: Desk Number: BSc and MSc Degree Examinations 2018-9 Department : BIOLOGY Title of Exam: Genetics Time Allowed: 1 Hour 30 Minutes Marking Scheme: Total marks available for
More informationHiPer 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 informationSYNTHETIC BIOLOGY AND THE HIGH SCHOOL CURRICULUM: LAB 1 _Lab_1
SYNTHETIC BIOLOGY AND THE HIGH SCHOOL CURRICULUM: LAB 1 http://openwetware.org/wiki/synthetic_biology_and_the_high_school_curriculum: _Lab_1 LAB 1: Eau that smell Comparing 2 competing designs to optimize
More informationSession 7 Glycerol Stocks & Sequencing Clones
Session 7 Glycerol Stocks & Sequencing Clones Learning Objective: In this lab you will prepare several of your clones for DNA sequencing and make glycerol stock cultures as a stable and uniform starting
More informationReading Lecture 3: 24-25, 45, Lecture 4: 66-71, Lecture 3. Vectors. Definition Properties Types. Transformation
Lecture 3 Reading Lecture 3: 24-25, 45, 55-66 Lecture 4: 66-71, 75-79 Vectors Definition Properties Types Transformation 56 VECTORS- Definition Vectors are carriers of a DNA fragment of interest Insert
More informationHOUR EXAM II BIOLOGY 422 FALL, In the spirit of the honor code, I pledge that I have neither given nor received help on this exam.
Name First Last PID Number - (Please Print) HOUR EXAM II BIOLOGY 422 FALL, 2007 In the spirit of the honor code, I pledge that I have neither given nor received help on this exam. 1 Signature 2 3 4 5 6
More informationIntroduction to pglo lab
Please take these notes carefully. You do not need to write anything in RED Introduction to pglo lab Bacteria Transformation What is a plasmid? A plasmid is a small circular piece of DNA (about 2,000 to
More informationExam Questions from Exam 2 Mutations, Bacterial Genetics, and Bacterial Gene Regulation
Exam Questions from Exam 2 Mutations, Bacterial Genetics, and Bacterial Gene Regulation 1. Drawn below is part of a wild-type gene. The DNA sequence shown encodes the last amino acids of a protein that
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 informationName 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 informationONTARIO 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 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 informationLecture 1. Basic Definitions and Nucleic Acids. Basic Definitions you should already know
Lecture 1. Basic Definitions and Nucleic Acids Basic Definitions you should already know apple DNA: Deoxyribonucleic Acid apple RNA: Ribonucleic Acid apple mrna: messenger RNA: contains the genetic information(coding
More informationArtificial Sweeteners Mutagenic Effects. Warren Austin Pittsburgh Central Catholic High School Second Year at PJAS
Artificial Sweeteners Mutagenic Effects Warren Austin Pittsburgh Central Catholic High School Second Year at PJAS Artificial Sweeteners Maybe derived from naturally occurring substances Excessive sweetness
More informationencodes 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 informationPacing/Teacher's Notes
Slide 1 / 31 New Jersey Center for Teaching and Learning Progressive Science Initiative This material is made freely available at www.njctl.org and is intended for the non-commercial use of students and
More informationSaccharomyces cerevisiae. haploid =
In this lecture we are going to consider experiments on yeast, a very useful organism for genetic study. Yeast is more properly known as Saccharomyces cerevisiae, which is the single-celled microbe used
More informationStudent 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 informationName_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 informationGenetic Background Page 1 PHAGE P22
Genetic Background Page 1 PHAGE P22 Growth of P22. P22 is a temperate phage that infects Salmonella by binding to the O-antigen, part of the lipopolysaccharide on the outer membrane. After infection, P22
More informationVirtual Laboratory Bacterial Transformation
Virtual Laboratory Bacterial Transformation Name: Before going to the Virtual lab, go to Bozeman site for AP Bio and watch: Lab 6 Molecular Biology *Note we have already done the Gel electrophoresis lab,
More informationpglo Transformation Lab Integrated Science 4 Redwood High School Name Per:
pglo Transformation Lab Integrated Science 4 Redwood High School Name Per: n Introduction To Transformation In this lab you will perform a procedure known as a genetic transformation. Remember that a gene
More informationBiol/Chem 475 Spring 2007
Biol/Chem 475 Spring 2007 Goal of lab: For most of the quarter, we will be exploring a gene family that was first discovered in fruitlfies and then found to be present in humans and worms and fish and
More informationTransformation of Bacillus subtilis with prit4501 and prit4502
Transformation of Bacillus subtilis with prit4501 and prit4502 Plasmids prit4501 and prit4502 were created by fusing the E. coli plasmid puc9 with the B. subtilis plasmid pbac. The recombinants therefore
More information# ml too many to count # ml 161/173 # ml 4/1
Biol 322 Fall 2012 Study Sheet for Quiz #2 Quiz #2 is scheduled for Thursday Nov 10 th and will be worth 30-40 pts. This quiz will cover: Mutagenesis Lab: Parts 1 & 2 Bacterial Genetics: F X F- Cross All
More informationThe yeast two-hybrid assay to discover if known proteins in the ethylene signaling pathway can physically interact with each other
The yeast two-hybrid assay to discover if known proteins in the ethylene signaling pathway can physically interact with each other Objective To perform the yeast two-hybrid assay, which is a powerful technique
More information7.03, 2005, Lecture 20 EUKARYOTIC GENES AND GENOMES I
7.03, 2005, Lecture 20 EUKARYOTIC GENES AND GENOMES I For the last several lectures we have been looking at how one can manipulate prokaryotic genomes and how prokaryotic genes are regulated. In the next
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 informationBacterial genetic exchange:transformation
Experiment 11 Laboratory to Biology III Diversity of Microorganisms / Wintersemester / page 1 Experiment Advisor Reading Objectives Background Literature www. Links Bacterial genetic exchange:transformation
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 informationLAB 1: Eau that smell
LAB 1: Eau that smell Compare 2 competing designs to optimize system performance Acknowledgements: This lab was developed with materials and guidance from the MIT 2006 igem team, as well as technical insights
More informationDNA Cloning with Cloning Vectors
Cloning Vectors A M I R A A. T. A L - H O S A R Y L E C T U R E R O F I N F E C T I O U S D I S E A S E S F A C U L T Y O F V E T. M E D I C I N E A S S I U T U N I V E R S I T Y - E G Y P T DNA Cloning
More information1a. What is the ratio of feathered to unfeathered shanks in the offspring of the above cross?
Problem Set 5 answers 1. Whether or not the shanks of chickens contains feathers is due to two independently assorting genes. Individuals have unfeathered shanks when they are homozygous for recessive
More informationSection B: The Genetics of Bacteria
CHAPTER 18 MICROBIAL MODELS: THE GENETICS OF VIRUSES AND BACTERIA Section B: The Genetics of Bacteria 1. The short generation span of bacteria helps them adapt to changing environments 2. Genetic recombination
More information7.03 Final Exam Review 12/19/2006
7.03 Final Exam Review 12/19/2006 1. You have been studying eye color mutations in Drosophila, which normally have red eyes. White eyes is a recessive mutant trait that is caused by w, a mutant allele
More informationLab Exercise: Examining Water Quality: Most Probable Number & Colilert Test Kit Lab
Lab Exercise: Examining Water Quality: Most Probable Number & Colilert Test Kit Lab OBJECTIVES 1. Understand the use of MPN to determine likely fecal water contamination. 2. Understand the use of MUG,
More informationTransforming E. Coli with pglo Plasmids
Name: Transforming E. Coli with pglo Plasmids AP Biology Transformation Background: Transformation is a process of transferring genetic information from one organism to another. In bacteria, a small circular
More information7.03 Final Exam. TA: Alex Bagley Alice Chi Dave Harris Max Juchheim Doug Mills Rishi Puram Bethany Redding Nate Young
7.03 Final Exam Name: TA: Alex Bagley Alice Chi Dave Harris Max Juchheim Doug Mills Rishi Puram Bethany Redding Nate Young Section time: There are 13 pages including this cover page Please write your name
More informationM. Dalbey/Bio 105M Isolation of E. coli - Isolation of E. coli from an Environmental Sample
Isolation of E. coli from an Environmental Sample We want to expand our horizons a bit beyond the domesticated lab strains of E. coli. In this exercise you will isolate "wild" E. coli strains from an environmental
More informationMCB 421 Second Exam October 27, 2004
MCB 421 Second Exam October 27, 2004 1. (10 pts) As discussed in class in complementation studies using F plasmids complementation can be confused with the products of homologous recombination between
More informationBiotechnology - Transformation Biology Concepts of Biology 7.1
Biotechnology - Transformation Biology 100 - Concepts of Biology 7.1 Name Instructor Lab Section Objectives: To gain a better understanding of: Use of Bacteria in Biotechnology DNA & Plasmid Structure
More informationUnderstanding the Cellular Mechanism of the Excess Microsporocytes I (EMSI) Gene. Andrew ElBardissi, The Pennsylvania State University
Understanding the Cellular Mechanism of the Excess Microsporocytes I (EMSI) Gene Andrew ElBardissi, The Pennsylvania State University Abstract: Hong Ma, The Pennsylvania State University The Excess Microsporocytes
More informationThe 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 informationBi 1x Spring 2016: LacI Titration
Bi 1x Spring 2016: LacI Titration 1 Overview In this experiment, you will measure the effect of various mutated LacI repressor ribosome binding sites in an E. coli cell by measuring the expression of a
More informationManipulating DNA. Nucleic acids are chemically different from other macromolecules such as proteins and carbohydrates.
Lesson Overview 14.3 Studying the Human Genome Nucleic acids are chemically different from other macromolecules such as proteins and carbohydrates. Nucleic acids are chemically different from other macromolecules
More informationWhat determines if a mutation is deleterious, neutral, or beneficial?
BIO 184 - PAL Problem Set Lecture 6 (Brooker Chapter 18) Mutations Section A. Types of mutations Define and give an example the following terms: allele; phenotype; genotype; Define and give an example
More informationBiological Sciences 50 Practice Final Exam. Allocate your time wisely.
NAME: Fall 2005 TF: Biological Sciences 50 Practice Final Exam A. Be sure to write your name on the top of each of page of the examination. B. Write each answer only on the same page as the pertinent question.
More informationHI-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 informationBacterial genetic exchange : Bacterial Transformation
Experiment 11 Laboratory to Biology III: Diversity of Microorganisms 1 Experiment 11 Bacterial genetic exchange : Bacterial Transformation Advisor Munti Yuhana myuhana@botinst.unizh.ch Textbook Chapters
More informationREGULATION 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 informationGenetics C Examination #3 - December 7, 1999
Genetics C3032 - Examination #3 - December 7, 1999 General instructions: Don't Panic. Be sure your name is on every page and that you write your exam in ink. Answer the questions in the space provided.
More informationMarch 15, Genetics_of_Viruses_and_Bacteria_p5.notebook. smallest viruses are smaller than ribosomes. A virulent phage (Lytic)
Genetics_of_Viruses_and_Bacteria_p5.notebook smallest viruses are smaller than ribosomes Adenovirus Tobacco mosaic virus Bacteriophage Influenza virus envelope is derived from the host cell The capsids
More informationChapter 9 Microbial Genetics
Chapter 9 Microbial Genetics You are expected to know details of 1) DNA replication 2) RNA synthesis (transcription) 3) Protein synthesis (translation) Genome & Genes A genome is all the genetic information
More informationCHAPTER 4A MAKING SURE YOU VE GOT A RECOMBINANT PLASMID. CHAPTER 4A STUDENT GUIDE 2013 Amgen Foundation. All rights reserved.
CHAPTER 4A MAKING SURE YOU VE GOT A RECOMBINANT PLASMID 55 INTRODUCTION When biologists clone a gene in order to produce human insulin, they create a recombinant plasmid that has the human insulin gene.
More informationPackaging of P22 DNA requires a pac site while packaging of lambda DNA requires a cos site. Briefly describe:
1). (12 Points) Packaging of P22 DNA requires a pac site while packaging of lambda DNA requires a cos site. Briefly describe: 1. The mechanisms used by P22 and lambda to package DNA. P22 uses a headfull
More informationTransduction of an Antibiotic Resistance Gene. Background
I Student Guide 21-1128 Name------------ Date Transduction of an Antibiotic Resistance Gene Background Transduction is a natural method of gene transfer that occurs in bacteria. The key player in transduction
More informationSafety Considerations. Notes for the last TWO exercises
Notes for the last TWO exercises! The notes that follow outline the activities for the last TWO exercises: 1- Molecular Biology (blue on your Calendar), and 2- Protein Expression (red on your Calendar).
More informationLab 2C: Basic Techniques. Dilute 10X TE Buffer to Make 1X TE Buffer Determine the Concentration of an Unknown DNA Sample Streak out bacteria colonies
Demonstration of sterile technique. Lab 2 Basic Techniques Lab 2A: Lab 2B: Lab 2C: Dilute 10X TE Buffer to Make 1X TE Buffer Determine the Concentration of an Unknown DNA Sample Streak out bacteria colonies
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 information