Recombinant DNA cloning

Chapter 13 Recombinant DNA Cloning and Creation of Chimeric Genes... 

Rat genome, 41 "Rational polytherapy," 233 Rawls, John, 80 Recombinant DNA cloning experiments with, 30 Cohen-Boyer patent on, 73 Recombination events, 57-58 Reeder, C. E., 229... 

Garrett, R. H., and Grisham, C. M. (1995). Recombinant DNA Cloning and Creation of Chimeric Genes. In Biochemistry. Orlando, FL Saunders. 

In order to manipulate or sequence a cloned DNA fragment. It first must be separated from the vector DNA. This can be accomplished by cutting the recombinant DNA clone with the same restriction enzyme used to produce the recombinant vectors originally. The cloned DNA and vector DNA then are subjected to gel electrophoresis, a powerful method for separating DNA molecules of different size. 

Bacteriophage Bacterial virus bacteriophage 1 is a biological reagent used in recombinant DNA cloning. 

Fig, 5. Amplified production of silicatein protein from recombinant DNA cloned in bacteria. DNA coding for the silicatein a protein was integrated enzymically into a double-stranded circular DNA vector molecule to produce a recombinant DNA. When introduced into bacteria, the recombinant DNA is replicated by intracellular enzymes to produce many copies in each cell. These DNA molecules serve as templates, directing the synthesis of the silicatein protein they encode. Rapid reproduction of the bacteria greatly amplifies this synthesis. 

Yet nucleic acids do it relatively quickly and with amazing precision. Indeed this reannealing (or hybridization) capability between a polynucleotide and a complementary piece of oligonucleotide (commonly called a printer or a probe) forms the basis of such diverse and critical techniques as dideoxynucleotide sequencing, site-specific mutagenesis, cDNA cloning, and hybridization screening of recombinant DNA clones. Nucleic acid hybridization also serves as the basis... 

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