Escherichia coli library construction

Sastry, L., Alting-Mees, M., Huse, W.D., Short, J.M., Sorge, J.A., Hay, B.N., Janda, K.D., Benkovic, S.J., and Lerner, R.A. (1989) Cloning of the immunological repertoire in Escherichia coli for generation of monoclonal catalytic antibodies Construction of a heavy chain variable region-specific cDNA library. Proc. Natl. Acad. Sci. USA 86, 5728-5732. 

Small peptides have also been obtained. Thus, the construction of a cosmid library in Escherichia coli, and cloning and heterogously expressing individual biosynthetic pathways, led to the discovery... 

An example of the former cloning strategy, outlined in Fig. 6.17, is provided by Rishi McManus (695), who have constructed a small, size-selected genomic DNA library in a plasmid vector of Escherichia coli using total DNA isolated from Echinococcus granulosus. Subsequent differential... 

The expression vector to be built in this demonstration will use the T5 promoter (5-8), which is constitutive in Escherichia coli. It can be made repressible by flanking it with lac operators, DNA sequences that bind to the lac repressor. To maximize the repression, we use an idealized perfectly symmetrical version of the operator (9) and space two operators around the promoter (10). Some of these elements (the T5 promoter and the lac operator) are present in the library of elements and can be added to the construct by dragging... 

As early as 1988 the poly-3-hydroxybutyrate (PHB) biosynthetic pathway from Alcaligenes eutrophus H16 has been cloned and expressed in Escherichia coli [269]. Initially, an A. eutrophus H16 genomic library was constructed by using cosmid... 

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