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- W3022892854 abstract "This chapter reviews the architecture of a gene construct suitable for high-level, regulated expression in E. coli and discusses the polymerase chain reaction based method for making overproducers. The construction of overproducing E. coli strains is a difficult and time-consuming task, primarily because of two factors: (1) the requirement that the gene or complementary DNA to be expressed be refashioned from a molecularly monoclonal source, and (2) the requirement to precisely assemble specific DNA sequences during construction of the recombinant gene. A polymerase chain reaction can overcome both these problems simultaneously by rapidly and selectively amplifying a target sequence from a mixed population, while at the same time allowing defined sequence alterations to be made in the amplification products. Ribosome-binding site (RBS) is fused directly to the upstream end of coding sequence. The promoter and transcription terminator are transcriptional control elements, they do not have to be positioned precisely with respect to the coding sequence, and their efficiency is essentially independent of the coding sequence; the RBS, on the other hand, is a translational control element, it must be positioned precisely with respect to the start codon, and varies in efficiency with different coding sequences." @default.
- W3022892854 created "2020-05-13" @default.
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- W3022892854 date "1995-01-01" @default.
- W3022892854 modified "2023-09-23" @default.
- W3022892854 title "Overproduction of Proteins Using Expression-Cassette Polymerase Chain Reaction" @default.
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- W3022892854 doi "https://doi.org/10.1016/b978-0-12-765561-1.50025-4" @default.
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