Volume 12, Issue 3 (11-1998)                   Med J Islam Repub Iran 1998 | Back to browse issues page

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AZARNOOSH M, ZEINALI S, TABATABAEI S, ZIAEE A. RT-PCR MEDIATED CLONING OF HUMAN GROWTH HORMONE GENE AND I TS EXPRESSION IN E. coli. Med J Islam Repub Iran 1998; 12 (3) :285-290
URL: http://mjiri.iums.ac.ir/article-1-1008-en.html
RT-PCR MEDIATED CLONING OF HUMAN GROWTH HORMONE GENE AND I TS EXPRESSION IN E. coli
M AZARNOOSH , S ZEINALI , SM TABATABAEI , A ZIAEE
From the Department of Biotechnology, Pasteur Institute of Iran, Tehran, Islamic Republic of Iran.
Abstract:   (4764 Views)
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.
Keywords: Human growth hormone, RT-PCR.
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Type of Study: Original Research: Basic Science in Medicine | Subject: Biological Sciences
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