Introduction Plasmids are circular‚ double stranded extrachromosomal DNA molecules that are found in bacteria which can self-replicate. They are naturally occurring DNA molecules advantageous to the host bacterium by carrying genes which specify metabolic capacities. (Garrett et al.‚ 2010) Besides‚ plasmids exist in a wide variety of sizes from a few thousands to hundreds of thousands of base pairs. Many plasmids have been engineered to serve as plasmids cloning vectors to carry genes. (Synder et
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E. Coli Transformation with Plasmid (pGal)‚ pGal Isolation‚ and Analysis of Plasmid DNA Felicia Osadi Bio 22 April 20‚ 2012 Transformation = group 10 Plasmid = group 7 RFLP = group 1 RESULTS Table I. Plasmid Transformation of E. Coli. Plate # | Agar plate | Type | Result | 1 | X-gal | Control | Extensive lawn growth | 2 | Ampr / X-gal | Control | Clear no bacterial growth | 3 | Ampr / X-gal | Transformation | 1 blue colony | Transformation efficiency = 1 transformants
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Electrophoresis Applying the concepts in the experiment to produce bands at the end of the Gel Electrophoresis stage Interpreting what these bands mean with accordance to how the plasmid was cleaved Methods and Materials: For the experiment we used several restriction endonucleases (BamHI‚ EcoRI‚ HindIII‚ PstI‚ ScaI‚ SaII)‚ ppBR322 plasmid DNA‚ TAE/TE Buffer‚ DNA Ladder (50 Bp)‚ Restriction Buffers‚ 1g of Agarose‚ 700ml of Distilled H2O. Equipment used for the experiment included: Agarose Gel Electrophoresis
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Insertion and amplification of EGFP protein into pET41a(+) Plasmid Introduction The overall purpose of the experiments conducted is to test the creation of recombinant plasmid using recombinant DNA technology. The research of recombinant DNA began with the use of E.Coli (Escherichia coli)‚ a common bacteria found in the intestines of warm-blooded or- ganisms (5). Scientists worked together and generated a way‚ from cloning and using recombi- nant research‚ to achieve recombinant DNA
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DIFFERENCES BETWEEN PLASMID AND CHROMOSOMAL DNA IN BACTERIA. Eukaryotes have two or more chromosomes‚ prokaryotes such as bacteria possess a single chromosome composed of double-stranded DNA in a loop. DNA is located in the nucleoid of the cell and is not associated with protein A plasmid is an extra-chromosomal DNA molecule separate from the chromosomal DNA which is capable of replicating independently of the chromosomal DNA. Plasmids usually occur naturally in bacteria. A chromosome
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Bacterial Transformation Bacteria and plasmid to produce Red Fluorescent Proteins Alejandra Lopez
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The purpose of this experiment was to create a new plasmid (pMV158GFP)‚ which lacks the malR protein and is mobile and has the GFP gene to be used to track the flow of genes between bacterial cells through conjugation. A plasmid with these qualities is necessary to create a plasmid that can be transferred to Gram-positive bacteria low in C&G (which are hard to transform with traditional means) by conjugation with other bacteria. Current vectors have the malR regulatory protein which imposes a problem
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are freezing and heat shocking the E. Coli bacteria that is then forced to take the plasmid DNA. The E. coli then transforms the pBLu plasmid‚ which carries the genes coding for two identifiable phenotypes. After following the Carolina Biological steps our lab worked well and we able to see some colonies of bacteria on the plates. The x-gal plate showed a significant amount of bacteria to confirm that the pBlu plasmid took over the E. coli strain. Introduction This lab was meant to reveal the variations
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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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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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