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[1] Recombinant DNA techniques

1979; Academic Press; Linguagem: Inglês

10.1016/0076-6879(79)68003-5

1557-7988

John F. Morrow,

Advanced biosensing and bioanalysis techniques

Publisher Summary This chapter discusses recombinant deoxyribonucleic acid (DNA) techniques. Recombinant DNA methods have become a mainstay of molecular genetics; they also contribute to solutions of practical problems, such as supplying hormones. A recombinant DNA method consists of joining DNA molecules in vitro and introducing them into living cells where they replicate. The essential ingredients of a recombinant DNA experiment include (1) a DNA vehicle (vector and replicon) that can replicate in living cells after foreign DNA is inserted into it, (2) a DNA molecule to be replicated (passenger) or a collection of them, (3) a method of joining the passenger to the vehicle, (4) a means of introducing the joined DNA molecule into a host organism in which it can replicate (DNA transformation or transfection), and (5) a means of screening or genetic selection for those cells that have replicated the desired recombinant molecule. Two mammalian hormones have been produced in bacteria by means of synthetic DNA. Polypeptides similar or identical to the ones found in eukaryotes have been synthesized in Escherichia coli (E. coli). By using recombinant DNA, the somatic recombination of immunoglobulin genes has been established and a large number of variable-region genes have been found. Intervening sequences (introns) have also been found in the DNA of eukaryotic cells.