The Over-Expression of Biologically Active Human Growth Hormone in a T5-Based System in Escherichia coli, Studying Temperature Effect

Author(s):

A. Zomorrodipour , B. Yakhchali , M. Khodabandeh , A. Deezagi , S.M. Hosseini Mazinani , S. Valian Borujeni , M. Ahmadzadeh Raji , M. Rahimi , H. Ahmadi Danesh , M.H. Sanati

Message:
Abstract:
We studied the expression of human growth hormone (hGH) in E. coli under a bacteriophage T5-base promoter in a pQE30 expression vector. For an efficient expression of hGH cDNA, a number of codons at the hGH N-terminal coding region were altered based on the E. coli major codons. An over-expression of hGH in the bacteria, carrying the recombinant plasmids, was observed at 37°C in the presence of IPTG. The over-expression was also observed at 30°C in the absence of IPTG. Therefore a temperature down-shift induction, 37°C to 30°C, was suggested to achieve an over-expression of recombinant hGH (rhGH) without the use of chemical inducers. The pQE30-hGH recombinant plasmids show high stability in the TG1 host in the non-selective conditions. In a batch fermentation condition, the purified rhGH was obtained with the yield of 53 mg/l of culture. We took advantage of the formation of inclusion bodies to recover the rhGH, followed by diafiltration and refolding steps. The purified rhGH was biologically active for its receptor-binding on IM9 cells.
Language:
English
Published:
Journal of Sciences, Islamic Republic of Iran, Volume:15 Issue: 1, Winter 2004
Page:
27
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