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| Abstract : Universiti Pendidikan Sultan Idris |
| Overexpression of a number of industrially- and medically-important proteins is always a challenge for most researchers as genes from bacterial hosts especially from the opportunistic pathogen, Pseudomonas aeruginosa, as well as from other Pseudomonads were reported to be poorly expressed in E. coli. Examples continue to be published demonstrating that certain Pseudomonas proteins cannot be expressed at satisfactory levels in E. coli owing to their promoter variations, absence of specific molecular activators and poor protein translation. Nevertheless, the extension of E. coli-based technology to Pseudomonas can be used to overcome such barriers. In this chapter, the evolution and modification of genetic tools ranging from types of promoters, elements for interspecies transfer, origins of replications, regulatory genes, genes facilitating in genome integration and host strains, used in the construction of Escherichia-Pseudomonas shuttle expression vectors will be explored and discussed in detail. Protein overexpression data as documented by the constructed shuttle expression vectors in this chapter, may offer a new strategy to express abundant amount of active proteins which can be an important determinant factor for successful nuclear magnetic resonance (NMR) and X-ray crystallography studies especially involving proteins from Pseudomonas and its related genus. ? 2021 by Nova Science Publishers, Inc. All rights reserved. |
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