ISOLATION AND CLONING OF PHYCOCYANIN ALPHA SUBUNIT GENE AND ITS PRODUCTION IN E.COLI EXPRESSION SYSTEM

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SHOJA Z.; RAJABI MEMARI H.; ROAYAEI ARDAKANI M.

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Journal: Journal of Molecular and Cellular Researches Year:2015 | Volume:28 | Issue:3 Page(s): 352-359

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Title

ISOLATION AND CLONING OF PHYCOCYANIN ALPHA SUBUNIT GENE AND ITS PRODUCTION IN E.COLI EXPRESSION SYSTEM

Author(s)

SHOJA Z. | RAJABI MEMARI H. | ROAYAEI ARDAKANI M. | Issue Writer Certificate

Keywords

CLONINGQ1

EXPRESSIONQ1

PHYCOCYANIN ALPHA SUBUNIT GENEQ2

SPIRULINA PLATENSISQ2

PET-43.1A++Q2

Abstract

Phycocyanin is a blue pigment in two eukaryote algal genera and in cyanobacteria as Spirulina. This pigment has various biology activities and are utilised in a number of applications in foods, cosmetics and pharmaceuticals. A lot advantage of phycocyanin studied by many researchers but the scale-up of these methods is difficult and expensive while production of recombinant phycocyanin is more convenience and inexpensive to scale up protein desire. The purpose of this study was to isolation and CLONING of PHYCOCYANIN ALPHA SUBUNIT GENE in EXPRESSION vector and production of recombinant protein in E.coli to provide industrial production of phycocyanin. The genomic DNA of SPIRULINA PLATENSISwas prepared and used for PCR as template. PHYCOCYANIN ALPHA SUBUNIT GENE amplified by designed specialize primers was cloned in a pET43.1a+ EXPRESSION vector, under the control of T7 promoter usingNdeI and NotI restriction enzymes. The CLONING of PHYCOCYANIN ALPHA SUBUNIT GENE is confirmed by colony PCR, digestion and DNA sequencing. The constructs were transformed in to E.coli strain BL21 (DE3). EXPRESSION of PHYCOCYANIN ALPHA SUBUNIT GENE was examined by 12.5 % SDS-PAGE analysis at 8 hrs after induction by IPTG. The SDS-PAGE analysis showed that alpha subunit phycocyanin was produced in E.coli EXPRESSION system. Our study provided the production of recombinant phycocyanin. Also overexpression of the synthetic alpha subunit phycocyanin in a bacterial system (E.coli BL-21) showed that E.colican be used to produce this desire protein in large quantity.

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APA: Copy

SHOJA, Z., RAJABI MEMARI, H., & ROAYAEI ARDAKANI, M.. (2015). ISOLATION AND CLONING OF PHYCOCYANIN ALPHA SUBUNIT GENE AND ITS PRODUCTION IN E.COLI EXPRESSION SYSTEM. JOURNAL OF MOLECULAR AND CELLULAR RESEARCH (IRANIAN JOURNAL OF BIOLOGY), 28(3), 352-359. SID. https://sid.ir/paper/248456/en

Vancouver: Copy

SHOJA Z., RAJABI MEMARI H., ROAYAEI ARDAKANI M.. ISOLATION AND CLONING OF PHYCOCYANIN ALPHA SUBUNIT GENE AND ITS PRODUCTION IN E.COLI EXPRESSION SYSTEM. JOURNAL OF MOLECULAR AND CELLULAR RESEARCH (IRANIAN JOURNAL OF BIOLOGY)[Internet]. 2015;28(3):352-359. Available from: https://sid.ir/paper/248456/en

IEEE: Copy

Z. SHOJA, H. RAJABI MEMARI, and M. ROAYAEI ARDAKANI, “ISOLATION AND CLONING OF PHYCOCYANIN ALPHA SUBUNIT GENE AND ITS PRODUCTION IN E.COLI EXPRESSION SYSTEM,” JOURNAL OF MOLECULAR AND CELLULAR RESEARCH (IRANIAN JOURNAL OF BIOLOGY), vol. 28, no. 3, pp. 352–359, 2015, [Online]. Available: https://sid.ir/paper/248456/en

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