INVESTIGATING OF THE EFFECT OF DECLINE IN GC CONTENT ON THE EXPRESSION OF HUMAN BASIC GROWTH FACTOR IN ESCHERICHIA COLI FIBROBLAST

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ALIBOLANDI M.; MIRZAHOSSEINI H.

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Paper Information

Journal: JOURNAL OF MICROBIAL BIOTECHNOLOGY Year:2009 | Volume:1 | Issue:2 (2) Page(s): 15-24

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Title

INVESTIGATING OF THE EFFECT OF DECLINE IN GC CONTENT ON THE EXPRESSION OF HUMAN BASIC GROWTH FACTOR IN ESCHERICHIA COLI FIBROBLAST

Author(s)

ALIBOLANDI M. | MIRZAHOSSEINI H. | Issue Writer Certificate

Keywords

GENE MODIFICATIONQ3

GC CONTENTQ3

RECOMBINANT PROTEIN EXPRESSIONQ3

HBFGFQ3

Abstract

Background and Objective: Translation initiation region (TIR) significantly affects the level of RECOMBINANT PROTEIN EXPRESSION in E .coli. In order to determine this effect we designed a long primer to modify TIR and reduce GC CONTENT.Materials and methods: We had pET-1006 (human basic Fibroblast Growth Factor cDNA inserted to pET-22b) as a template .Site directed mutagenesis were applied to alter the 5′ end of HBFGF cDNA and decrease GC CONTENT through nucleotide changes, without changing amino acid sequences .The amount of produced HBFGF expressions by mutant and wild gene was quantified based on SDS-PAGE densitometry assay, western blotting and ELISA.Results: Obtained result proved that expression rate of mutant gene was increased when compared to wild type. This serious difference might relate to the mRNA efficiency for translation. Conclusion: It shows that reduction of GC CONTENT will decrease the stability of secondary structure of mRNA and cause enhancement in translation.

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APA: Copy

ALIBOLANDI, M., & MIRZAHOSSEINI, H.. (2009). INVESTIGATING OF THE EFFECT OF DECLINE IN GC CONTENT ON THE EXPRESSION OF HUMAN BASIC GROWTH FACTOR IN ESCHERICHIA COLI FIBROBLAST. JOURNAL OF MICROBIAL BIOTECHNOLOGY, 1(2 (2)), 15-24. SID. https://sid.ir/paper/181407/en

Vancouver: Copy

ALIBOLANDI M., MIRZAHOSSEINI H.. INVESTIGATING OF THE EFFECT OF DECLINE IN GC CONTENT ON THE EXPRESSION OF HUMAN BASIC GROWTH FACTOR IN ESCHERICHIA COLI FIBROBLAST. JOURNAL OF MICROBIAL BIOTECHNOLOGY[Internet]. 2009;1(2 (2)):15-24. Available from: https://sid.ir/paper/181407/en

IEEE: Copy

M. ALIBOLANDI, and H. MIRZAHOSSEINI, “INVESTIGATING OF THE EFFECT OF DECLINE IN GC CONTENT ON THE EXPRESSION OF HUMAN BASIC GROWTH FACTOR IN ESCHERICHIA COLI FIBROBLAST,” JOURNAL OF MICROBIAL BIOTECHNOLOGY, vol. 1, no. 2 (2), pp. 15–24, 2009, [Online]. Available: https://sid.ir/paper/181407/en

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