PCR and cloning of recombinant growth hormone cDNA from Cholistani cow

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Research Paper 01/11/2019
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PCR and cloning of recombinant growth hormone cDNA from Cholistani cow

Maryam Arshad, Tahira Ruby, Mirza Imran Shahzad, Gulnaz Afzal, Muhammad Rafay
Int. J. Biosci.15( 5), 382-386, November 2019.
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Cholistani cow breed is a zebu (Bos indicus) or one humped breed of cattle being reared by the nomadic pastoralists of Cholistan desert, Pakistan. With an increasing human population and improving living standards, the demand for meat and milk is also increasing. To cope with such conditions in Pakistan, especially in Cholistan, recombinant technology is used. Importance of recombinant growth hormone rGH in milk and meat production bovine growth hormone bGH has been cloned from a number of species like buffalo, sheep, goat, sheep, and horse etc. The present study was carried out to clone bGH from Chlostani cow. For this purpose, total cellular RNA was extracted from the pituitary gland of freshly slaughtered animal, cDNAs were synthesized and amplified with the help of sequence specific primers. The amplified products were confirmed through restriction digestion and finally cloned in Thyamine Adenine T/A cloning vector followed by transformation in E. coli strain DH5ɑ. Positive clones were confirmed by restriction digestion and sequence analysis confirmed clones were stored at -70˚C as glycerol stock. By using cDNA growth hormone, milk production in young cows can be enhanced. More quantity of milk will compensate the needs of growing population.


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