molecules. In this exercise you will attempt to transform E. coli with a plasmid containing genes for phosphorescence‚ and for resistance to the antibiotic ampicillin. Plasmids are usually small‚ circular pieces of DNA that replicate independently of the bacterial chromosome. Many plasmids have been modified to function as vectors‚ or vehicles for transferring genes of interest from one organism to another. Plasmids that have been modified as cloning vectors usually possess a selectable marker
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Electrophoresis Christina Qi 2/16/07 Aim: How can a plasmid be engineered to include a foreign piece of DNA and how does gel electrophoresis separate DNA molecules present in a mixture? Hypothesis: If the pGLO plasmid is inserted into competent Escherichia coli cells‚ then the transformed bacteria will be resistant to ampicillin and will glow green under UV light. If samples of DNA are cut using certain restriction enxymes and separated using gel electrophoresis‚ then the smaller
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1. Plasmids Plasmids are closed circular‚ double-stranded‚ extrachromosomal DNA molecules which occur naturally in bacteria‚ yeast‚ and some higher eukaryotic cells‚ and exist in a parasitic or symbiotic relationship with their host cell (Lodish et al.‚ 2000) The main application of plasmids is as cloning vectors in gene cloning. In gene cloning‚ a fragment of DNA‚ containing the gene to be cloned is inserted into a circular molecule called the “vector” to produce recombinant DNA molecule. Plasmids
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Manipulation Chapter 1 Chapter 2 Chapter 3 Chapter 4 Chapter 5 Chapter 6 Chapter 7 Chapter 8 Chapter 9 Chapter 10 Gene manipulation: an all-embracing technique Basic techniques - (POGC02.pdf‚ 1‚560KB) Cutting and joining DNA molecules Basic biology of plasmid and phage vectors Cosmids‚ phasmids and other advanced vectors Cloning strategies Additional updated information on Cloning strategies Sequencing and mutagenesis Cloning in bacteria other than E. coli Cloning in Saccharomyces cerevisiae and other
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AP ESSAY ANSWERS: 16-20 1. Information transfer is fundamental to all living organisms. For TWO of the following examples‚ explain in detail‚ how the transfer of information is accomplished. A) The genetic material in one eukaryotic cell is copied and distributed to two identical daughter cells. B) A gene in a eukaryotic cell is transcribed and translated to produce a protein. C) The genetic material from one bacterial cell enters another via transformation‚ transduction or conjugation
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molecule in Figure 20.1? A) ligase B) transcriptase C) a restriction enzyme D) RNA polymerase E) DNA polymerase Answer: C Topic: Concept 20.1 Skill: Application/Analysis 2) Assume that you are trying to insert a gene into a plasmid. Someone gives you a preparation of genomic DNA that has been cut with restriction enzyme X. The gene you wish to insert has sites on both ends for cutting by restriction enzyme Y. You have a plasmid with a single site for Y‚ but not for X. Your strategy should
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rDNA amplicon with Hinf1 on 16s rDNA strains (II) The amplification and cloning of a specific stress gene given by * Designing primers for cloning * cloning stress gene into cloning vector (pGEM-T) * cloning stress gene into expression plasmid(pET23b) – not done Literature Review Lactic acid bacteria (LAB) LAB are defined as bacteria that produce lactic acid as their major fermentation production. The examples of LAB include Streptococcus‚ Enterococcus‚ Leuconostoc and Lactococcus
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4) EcoRI is a restriction enzyme that is used widely by scientists. EcoRi specifically cuts DNA containing the sequence GAATTC. The ends of the DNA fragments created by ECORI are called sticky ends because they contain single-stranded DNA that is complementary. 5) An
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genetic analysis as well as practical applications in medicine‚ agriculture‚ and industry. C. Fundamental changes in our society are occurring as a result of genetic engineering. II. Making recombinant DNA Overview: Isolate DNA à Cut with restriction enzymes à Ligate into cloning vector à transform recombinant DNA molecule into host cell à each transformed cell will divide many‚ many times to form a colony of millions of cells‚ each of which carries the recombinant DNA molecule (DNA clone)
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Module 1 |Long answers 1 | |Question 1 | |Question 2 | |Question 4
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