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Lab Report Of The Experiment Of Conjugation Of E. Coli

2244 words - 9 pages

Abstract:Conjugation is a natural occurring process that involves the transfer of DNA from one cell into another through a physical connection between the cells. In the following experiment, two strains of Escherichia coli bacterial cells (donor F'lac+strs and recipient F-lac-strr) underwent conjugation to produce a transconjugant strain (F'lac+strr). MAC plates and streptomycin were utilized to determine if conjugation had occurred. When plated, the donor colonies appeared red and the recipient colonies appeared white. The transconjugant plates showed red and white colonies. Using alkaline lysis miniprep, a DNA plasmid was isolated from the donor and transconjugant strains and FIGE electrophoresis was used to determine the size of the plasmid. The conjugation efficiency was found to be 16.25% and the plasmid DNA was approximately 97 kilobases long. The results show that the F' plasmid was effectively transferred from the donor cells into the recipient cells via conjugation.Introduction:Bacterial conjugation is the unidirectional transfer of either genomic DNA or plasmid DNA from a donor bacterial cell to a recipient bacterial cell by cell-to-cell contact via a sex pilus (Snustad & Simmons, 2006). Conjugation was first discovered by Lederberg and Tatum in 1946. In their experiment, they grew two strains of bacteria in separate vessels with rich medium and then together in one vessel containing the same medium. Then, they spread the three vessel contents onto medium agar plates and incubated them overnight at 37˚C. The only plate that showed cell growth was the plate containing the mixture of the two bacterial strains. The other two plates showed no growth. This experiment proved that in order for recombination to occur, the two strains must come in contact with one another (Lederberg, Tatum, 1946).In 1950, Bernard Davis discovered that cell-to-cell contact was required to obtain a transconjugant. Using a U tube containing a sintered filter between the two sides of the tube, he added two types of bacteria (donor and recipient) to each side of the tube. Because of the filter, Davis never observed conjugation. This further proved that in order for conjugation to occur, the cells must come into physical contact.In order for cells to undergo conjugation, one cell must contain a fertility factor (F). William Hayes discovered this F factor in 1952. The F factor, which is a small circular molecule of DNA (plasmid), controls the synthesis of F pili that connect donor and recipient cells during conjugation. These F factors are approximately 105 basepairs in size.In bacterial conjugation, a donor cell containing the F plasmid is referred to as an F+ cell while a recipient cell that lacks the plasmid is an F- cell. When an F+ cell mates with an F- cell (conjugation), the F+ plasmid is transferred. Both the donor and recipient cells become F+ cells and contain the F plasmid. While transferring the F+ plasmid, sometimes the plasmid is integrated into...

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