A gene cloning

Xia, Z.-J. et al. (2008) Discovery of the autonomously replicating plasmid pMFl from Myxococcus fulvus and development of a gene cloning system in Myxococcus xanthus. Appl Environ. Microbiol, 74 (7), 1980-1987. 

Matsushima P, Balti RH. A gene cloning system for Streptomyces tovocoensis. Microbiology 1996 142 261-267-... 

Hasegawa, M. A gene cloning system in Micromonospom which revealed the organization of biosynthetic genes of fortimicin A (astromicin). Actinomycetol 1991 5 126-131. 

Figure 1. Schematic representation of basic steps involved in a gene cloning procedure, cat, chloramphenicol transacetylase.

Clone Genetically identical copy (of a gene, cell or organism)... 

The basic steps of gene cloning first involve cutting a precise DNA segment (gene) from a donor source DNA by use of a restriction enzyme (Figure 45.2). At the same time, a small looped... 

DNA from a gene contains hundreds to thousands of nucleotide units for which the sequence is needed in order to interpret its code. Sequencing methods require only small amounts (5 (tg) of purified DNA, which can be produced by cloning. Automated sequencers are available that can daily sequence DNA containing hundreds of nucleotide units. 

Gene cloning is a method that uses recombinant technology to insert a gene into a vector DNA (plasmid obtained from a bacterium). The modified vector is put back into the bacterium, which then reproduces endlessly (clones) the new gene as well as the others in the vector. 

After a desired clone is obtained and mapped with restriction enzymes, further analysis usually depends on the deterrnination of its nucleotide sequence. The nucleotide sequence of a new gene often provides clues to its function and the stmcture of the gene product. Additionally, the DNA sequence of a gene provides a guidepost for further manipulation of the sequence, for example, lea ding to the production of a recombinant protein in bacteria. 

Bolivar, F., Rodriguez, R. L., Greene, P. J., et al., 1977. Construction and characterization of new cloning vehicles. II. A multipurpose cloning system. Gene 2 95-113. A paper describing one of the early plasmid cloning systems. 

Clone, identical cells (with respect to genetic constitution) derived from a single cell by asexual reproduction. Receptors can be cloned into cells by inserting a gene into the cell line a colony of cells results that are identical and all have the expressed receptor. 

Lee, D. H., et al. (1993). Molecular cloning and genomic organization of a gene for luciferin-binding protein from the dinoflagellate Gonyaulax polyedra. J. Biol. Chem. 268 8842-8850. 

Abstract In 2007, the world celebrated the 50th anniversary of the discovery of interferon (IFN) by Isaacs and Lindemnann. Subsequently, the IFN-a gene was cloned, fully sequenced and IFN-a was produced in recombinant form. Recombinant IFN-a is now used as the basis for treatment of chronic hepatitis C virus infection and can also be used to treat certain forms of chronic hepatitis B virus infections. IFNs have also been used in other viral infections, although with less success. The antiviral mechanisms of IFNs are reviewed in this chapter as well as the utility of IFNs in the treatment of persistent viral infections. 

---