rt-pcr mediated cloning of human growth hormone gene and i ts expression in e. coli

Authors

m azarnoosh from the department of biotechnology, pasteur institute of iran, tehran, islamic republic of iran.

s zeinali

sm tabatabaei

a ziaee

abstract

human growth hormone (hgh) genomic sequence containing 5 exons and 4 introns was cloned in pcdna-3 and the constructed plasmid was subsequently used for transfection ofnlli-3t3 cell line using lipofection technique. expression of hgh in stably transfected cells was assayed using elisa. total rna was extracted from transfected cells and hgh cdna was amplified by rt-pcr using specific primers. the product thus obtained was cloned in pgem-t vector and the presence of the growth hormone coding region was verified by restriction enzyme analysis and southern blotting. the hgh cdna fragment was cloned in pqe-30 and the expression of hgh gene in e. coli was assayed using elisa and immunoblotting. in this experiment 20.9 µg/ml purified rhgh was obtained.

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Journal title:
medical journal of islamic republic of iran

جلد ۱۲، شماره ۳، صفحات ۲۸۵-۲۹۰

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