1. [25] Bacterial expression vectors for calmodulin
- Author
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Anthony R. Means, John A. Putkey, and Patricia V. Donnelly
- Subjects
Genetics ,chemistry.chemical_compound ,Expression vector ,Glycosylation ,Plasmid ,chemistry ,Complementary DNA ,Vector (molecular biology) ,Biology ,Gene ,Fusion protein ,DNA-binding protein ,Cell biology - Abstract
Publisher Summary This chapter discusses expression plasmids that are designed to utilize a cDNA for the production of an unfused protein product. Recently, several groups applied bacterial expression systems to the study of Ca 2+ -binding proteins. Calcium-binding properties of the sea urchin spec proteins were demonstrated by an evaluation of fusion proteins produced in bacteria. A bacterial expression system is usually the method of choice for production of proteins that do not require extensive posttranslational modifications, such as glycosylation, for their activity or function. Bacterial expression vectors can produce either fused or unfused proteins. Fusion proteins are common products of expression libraries that use vectors such as the phage λgt11 or the plasmid pUC8, where the foreign protein is expressed as part of the β-galactosidase-selectable marker. The easiest approach to consider for constructing an expression plasmid is to select an existing vector that allows insertion of the desired gene in an appropriate position relative to the promoter. Construction of any expression plasmid requires a good strategy and familiarity with basic manipulations of DNA.
- Published
- 1987
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