Generation of new gene constructs for expression of ostrich growth hormone gene in prokaryotic and eukaryotic systems
Subject Areas : Microbial BiotechnologyHossein Fathpour 1 , Abbas Doosti 2 , Maryam Ghasemi 3 , Hamidreza Kabiri 4
1 - Associate Professor, Biotechnology Research Center, Islamic Azad University, Shahrekord Branch, Shahrekord, Iran
2 - Assistant Professor, Biotechnology Research Center, Islamic Azad University, Shahrekord Branch, Shahrekord, Iran.
3 - MS.c., Biotechnology Research Center, Islamic Azad University, Shahrekord Branch, Shahrekord, Iran.
4 - BS.c., Biotechnology Research Center, Islamic Azad University, Shahrekord Branch, Shahrekord, Iran.
Keywords: Cloning, Ostrich, Growth hormone (GH),
Abstract :
Background & Objectives: Growth of ostrich depends to employment of growth hormone and proper nutrition. The ostrich growth hormone is a polypeptide that stimulates growth and cell reproduction. The aim of this study was to clone and generation of two new gene constructs for expression of ostrich growth hormone gene in prokaryotic and eukaryotic systems. Materials & Methods: In this study, RNA was extracted from ostrich pituitary gland and cDNAs synthesized using RT-PCR method. These cDNAs were cloned into TOPO vector based on T/A method. Then, cDNA sub-clones of ostrich growth hormone were prepared on pcDNA3.1 and pET32 vectors. Results: Cloning of ostrich growth hormone cDNA in TOPO vector was confirmed by PCR and endonuclease restriction digest. After sub-cloning of this cDNA, pcDNA3.1-gh and pET32-gh recombinant vectors were generated and the presence of genes were confirmed using restriction digestion. Conclusion: According to the results, the new recombinant vectors generated in this study can serve as an effective approach for production of the ostrich growth hormone in commercial levels.