the over-expression of biologically active human growth hormone in a t5-based system in escherichia coli, studying temperature effect
نویسندگان
چکیده
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.
منابع مشابه
The Over-Expression of Biologically Active Human Growth Hormone in a T5-Based System in Escherichia coli, Studying Temperature Effect
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...
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عنوان ژورنال:
journal of sciences islamic republic of iranجلد ۱۵، شماره ۱، صفحات ۰-۰
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