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Recombinant Dna Lab Report

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Preparation of Recombinant Intermediates; Topologically different forms of DNA

INTRODUCTION
The gene is the cornerstone of the Molecular Biology techniques. These genes can be isolated and amplified for the better study. One of the most important methods in Molecular Biology is the insertion of desired gene or gene of interest into a vehicle or vector that can be replicated in living cells. This process is called cloning. The result of these two DNAs combination is called recombinant DNA molecule. Crossing-over which is a genetic process will produce recombinant DNA normally. This term is given for the DNA molecule produced by joining the segments derived from different biological sources. Host cell is the one which is the place to keep these recombinant DNA molecule and is prokaryotic or eukaryotic (mostly prokaryotic). When the host cell replicates, the piece of DNA within the vector will be get replicated. Thus, got the amplified foreign DNA, which is numerous in number and can be purified and for the further analysis (Kary B Mullis, 1990)
Geneticists use recombination term to describe the outcome of crossing over …show more content…

There are three different bacterial systems present. RecE, RecF, and RecBCD are the bacterial systems. The RecBCD is most important in bacteria. RecBCD enzyme will initiate the recombination. Enzyme is also called as exonuclease V, which have both nuclease and helicase activity. The enzyme complex have three subunits; RecB, RecC, and RecD. It is a bipolar DNA helicase enzyme. RecB and RecD have ATP-dependant helicase activity. RecB travel on 3'→5' direction on one strand and RecD on the other strand on 5'→3' direction. RecC recognizes specific sequence in DNA, called chi (Crossover Hotspot Instigator) site (Pranav Kumar and Usha Mina,

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