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Information Journal Paper

Title

CONSTRUCTION OF NEW GENETIC TOOLS AS ALTERNATIVES FOR PROTEIN OVEREXPRESSION IN ESCHERICHIA COLI AND PSEUDOMONAS AERUGINOSA

Author(s)

 WONG CHEE FAH | ZALIHA RAJA ABD. RAHMAN RAJA NOOR | BASRI MAHIRAN | SALLEH ABU BAKAR | Issue Writer Certificate 

Pages

  194-200

Keywords

T7(A1/O4/O3) 

Abstract

 Background: Pseudomonas protein expression in E. coli is known to be a setback due to signifi cant genetic variation and absence of several genetic elements inE. coli for REGULATION and activation of Pseudomonas proteins. Modifi cations in promoter/repressor system and shuttle plasmid maintenance have made the expression of stable and active Pseudomonas protein possible in both Pseudomonas sp. and E. coli.Objectives: Construction of shuttle expression vectors for REGULATION and OVEREXPRESSION of Pseudomonas proteins in Pseudomonassp. and E. coli.Materials and Methods: Pseudomonas-Escherichia shuttle expression vectors, pCon2 (3), pCon2 (3) -Kan and pCon2 (3) -Zeo as well asE. coli expression vectors of pCon4 and pCon5 were constructed from pUCP19-, pSS213-, pSTBlue-1- and pPICZαAbased vectors. Protein OVEREXPRESSION was measured using ELASTASE STRAIN K as passenger enzyme in elastino lytic activity assay.Results: The integration of two series of IPTG inducible expression cassettes in pCon2 (3), pCon2 (3) -Kan and pCon2 (3) -Zeo, each carrying anE. coli lac-operon based promoter, Plac, and a tightly regulated T7 (A1/O4/O3) promoter/repressor system was performed to facilitate OVEREXPRESSION study of the organic solvent-tolerant ELASTASE STRAIN K. These constructs have demonstrated an elastino lytic fold of as high as 1464.4 % in comparison to other published constructs. pCon4 and pCon5, on the other hand, are series of pCon2 (3) -derived vectors harboring expression cassettes controlled byPT7 (A1/O4/O3) promoter, which conferred tight REGULATION and repression of basal expression due to existence of respective double operator sites, O3 andO4, and LACIQ.Conclusions: The constructs off ered remarkable assistance for OVEREXPRESSION of heterogeneous genes in Pseudomonas sp. andE. coli for downstream applications such as in industries and structural biology study.

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    APA: Copy

    WONG, CHEE FAH, ZALIHA RAJA ABD. RAHMAN, RAJA NOOR, BASRI, MAHIRAN, & SALLEH, ABU BAKAR. (2017). CONSTRUCTION OF NEW GENETIC TOOLS AS ALTERNATIVES FOR PROTEIN OVEREXPRESSION IN ESCHERICHIA COLI AND PSEUDOMONAS AERUGINOSA. IRANIAN JOURNAL OF BIOTECHNOLOGY, 15(3), 194-200. SID. https://sid.ir/paper/706086/en

    Vancouver: Copy

    WONG CHEE FAH, ZALIHA RAJA ABD. RAHMAN RAJA NOOR, BASRI MAHIRAN, SALLEH ABU BAKAR. CONSTRUCTION OF NEW GENETIC TOOLS AS ALTERNATIVES FOR PROTEIN OVEREXPRESSION IN ESCHERICHIA COLI AND PSEUDOMONAS AERUGINOSA. IRANIAN JOURNAL OF BIOTECHNOLOGY[Internet]. 2017;15(3):194-200. Available from: https://sid.ir/paper/706086/en

    IEEE: Copy

    CHEE FAH WONG, RAJA NOOR ZALIHA RAJA ABD. RAHMAN, MAHIRAN BASRI, and ABU BAKAR SALLEH, “CONSTRUCTION OF NEW GENETIC TOOLS AS ALTERNATIVES FOR PROTEIN OVEREXPRESSION IN ESCHERICHIA COLI AND PSEUDOMONAS AERUGINOSA,” IRANIAN JOURNAL OF BIOTECHNOLOGY, vol. 15, no. 3, pp. 194–200, 2017, [Online]. Available: https://sid.ir/paper/706086/en

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