Recombinant plasmid DNA

Most of the known microorganisms to be active for BDS (by the date that patent [408] was introduced), were considered in this invention. Furthermore, their mutational or engineered derivatives, enzymes, cell-free extracts, recombinant enzymes, recombinant DNA, plasmids, vectors, and fragments were also incorporated in the intellectual property document. The mentioned operating conditions regard ambient temperature, mechanical agitation and a 1 9 biocatalyst/petroleum ratio. 

Recombinant DNA plasmids had no significant effect on survival of recombinant strains. 

Figure 10.9 Non-PCR random mutagenesis. Overview of a random site-directed mutagenesis method. Obtain template DNA [ssDNA (—) strand] Step 1 anneal spiked oligodeoxy nucleotide mutagenic primer (blue) with desired mutation range Step 2 extend and incorporate mutagenic primer into new (-f) strand containing dCTPaS Step 3 digest the template DNA (—) strand with Nci I and ExoIII, then reynthesize (—) strand to include mutations into (—) strand as well Step 4 transform E. coli with random mutant recombinant DNA plasmid ready for selection, DNA purification and sequence identification of mutant recombinant DNAs.

Bacterial cells are bathed in a solution containing recombinant DNA plasmids, which diffuse into the cells. When the bacteria reproduce, they replicate all the genes, including the recombinant DNA plasmid. Bacteria multiply quickly, and soon a large number of... 

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