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Original Research

Egypt. J. Exp. Biol. (Bot.). 2013; 9(1): 55-63


CLONING AND EXPRESSION OF BOVINE SOMATOTROPIN GROWTH HORMONE GENE FROM LOCAL COWS BREEDS OF BASRAH CITY, IRAQ

Mohamad A. Maarich AL-hajaj Othman R. Monther.




Abstract

The present study, have undertaken the cloning of bovine growth hormone or bovine somatotropin gene from local cows breed in Basrah city, and optimizing the conditions for its high-level expression in Escherichia coli. To achieve this goal the cDNA for bovine growth hormone was inserted in to the Pst1 site of pBR322 via the poly dC: poly dG joining technique to produce the recombinant vector. The cloning vector was then transformed into E. coli HB101. The efficiency of transformation also determined to be 2 × 107 cfu/µg. The fermentation strategy for high cells density growth of E. coli harboring the bGH gene was carried out and the highest cells density level was determined to be 1.540 ¬¬¬at OD600 after 8 hs of cells growing. Following solubilization and refolding, bovine growth hormone was purified by using ion exchange chromatography and the result was a single visible band with 22KDa on 12% SDS-Polyachrylamid gel which represents the purified bGH.

Key words: Bovine Growth hormone, Recombinant bovine somatotropin, milk, cow reproduction






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