Recombinant DNA libraries

L3. Lichter, P., Cremer, T., Borden, J., Manuelidis, L., and Ward, D. C., Delineation of individual human chromosomes in metaphase and interphase cells by in situ suppression hybridization using recombinant DNA libraries. Hum. Genet. 80, 224-234 (1988). 

Brody, Griffith J, Cuticchia AJ, Arnold J, Timberlake WE. Chromosome-specific recombinant DNA libraries from the fungus Aspergillus nidulans. Nucleic Acids Res 19 3105 3109, 1991. 

EPO is present in serum and (at very low concentrations) in urine, particularly of anaemic individuals. This cytokine/hormone was first purified in 1971 from the plasma of anaemic sheep, and small quantities of human EPO were later purified (in 1977) from over 2500 1 of urine collected from anaemic patients. Large-scale purification from native sources was thus impractical. The isolation (in 1985) of the human EPO gene from a genomic DNA library facilitated its transfection into CHO cells. This now facilitates large-scale commercial production of the recombinant human product (rhEPO), which has found widespread medical application. 

The diversity of antibodies can be increased further with phage display libraries and recombinant DNA technology. In recombinant DNA... 

Workers in the early 1970s recognized that restriction enzymes provided tools not only for DNA mapping but also for construction of new DNA species not found in nature. A collection of recombinant DNA species consisting of many passenger sequences joined to identical vector molecules is called a library. Individual recombinant DNAs are isolated from single clones of the library for detailed analysis and manipulation. 

Catalytic antibodies, like enzymes, must be isolated and purified to homogeneity before they can be studied. Initially this was done by using the hybridoma technique for isolation of monoclonal antibodies (Box 31-A). After induction of antibody formation by injecting a selected hapten into a mouse, large numbers of monoclonal antibodies had to be tested for catalytic activity. Even if several thousand different monoclonal antibodies were tested, only a few with catalytic properties could be found.1 Newer methods have incorporated recombinant DNA techniques (Box 31-A) and use of combinatorial libraries and phage display.) Incorporation of acidic or basic groups into the haptens used to induce antibody formation may yield antibodies capable of mimicking the acid-base catalysis employed by natural enzymes. 0... 

According to the opponents, the main argument against granting a patent was the lack of novelty of the recombinant DNA sequences claimed, which included the DNA sequences that coded interferon-alpha (IFN-alpha) as an insert. This prior art referred to DNA genome libraries, which are fractioned segments (15-20 Kb) of chromosomal DNA of human fetuses, cloned as derivatives of recombinant X phages. 

Construction of a mAb combinatory library expressed in a bacterial system (from Recombinant DNA, 2/e by James D. Watson, et al. 1992 by James D. Watson, Michael Gilman, Jan Wirkowski, and Mark Zoller. Used with permission). 

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