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Chapter 27 : Plasmid Vectors for Gene Cloning and Expression

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Abstract:

While plasmid vectors were initially designed for gene cloning and DNA analysis in , shuttle vectors for gene transfer between and other model organisms for gene function analysis and protein production were quickly developed. In an attempt to shed some light on the future development and utilization of plasmid vectors, this chapter summarizes some of the historical events in plasmid vector development and provides some critical considerations of the various types of vectors for gene cloning and gene expression. Gene expression is a complex process that involves dynamic steps that can be regulated at multiple levels, which include transcriptional (transcription initiation, elongation, and termination), posttranscriptional (RNA splicing, RNA translocation, RNA stability), translational (translation initiation, elongation, and termination), and posttranslational (protein splicing, translocation, stability, and modifications). A few general considerations may be used to achieve the desired level and duration of gene expression for a particular task. They are: (i) gene copy number and plasmid stability; (ii) transcription rate, inducibility, and RNA stability; (iii) translation efficiency and protein stability; (iv) protein solubility, functionality, and downstream utility; (v) gene delivery efficiency and protein targeting; (vi) host cell engineering; and (vii) cell growth condition and optimization. A number of approaches have been routinely used to generate a diversified pool of a target gene or a gene family. These approaches include random mutagenesis, exon swap, gene shuffling, and molecular evolution.

Citation: Lu Q. 2004. Plasmid Vectors for Gene Cloning and Expression, p 545-566. In Funnell B, Phillips G (ed), Plasmid Biology. ASM Press, Washington, DC. doi: 10.1128/9781555817732.ch27

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Tables

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Table 1

Major commercial suppliers of plasmid vectors for gene cloning and expression

Citation: Lu Q. 2004. Plasmid Vectors for Gene Cloning and Expression, p 545-566. In Funnell B, Phillips G (ed), Plasmid Biology. ASM Press, Washington, DC. doi: 10.1128/9781555817732.ch27