Plasmid Based Expression and Bioactivity Evaluation of Caprine Growth Hormone Gene Cloned from a Local Pakistani Goat Breed Beetal

• Published in: International journal of agriculture and biology 2014-06, Vol.16 (3)
• Main Authors: Hamama Islam, Butt Mirza, Shahzad, Imran, Bashir, Qudsia, Arif, Muhammad, Nadeem Saqib, Khanum, Azra
• Format: Article
• Language: English
• Subjects: Capra hircus
• ISSN: 1560-8530; 1814-9596

Growth hormone cDNA of Beetal goat (Capra hircus) an indigenous breed of Punjab Pakistan was amplified by RT PCR and gene including leader sequence was cloned in pTZR57 cloning vector. The cGH-pTZR57 clone was confirmed by restriction digestion and sequence analyses before finally sub-cloning the gene in pND - a mammalian expression vector. The clones were again confirmed by restriction digestion and PCR analyses. Highly purified super coiled recombinant caprine growth hormone-pND (rcGH-pND) construct was used to transfect Vero cell lines for expression studies. The in vitro expression of cGH was determined by dot-ELISA. After confirming its in vitro cell line based expression rcGH-pN pND constructs and phosphate buffer saline (PBS) were separately injected to 4 weeks old balb/c female mice intramuscularly. Total forty five animals were used to determine the biological activity and randomly divided in to three groups with fifteen mice in each group. Five animals from each group were used to monitor the in vivo biological activity by evaluating the body weight gain and tibia epiphyseal width assays from zero week up to four weeks with one week gap. Significant increase (P less than 0.05) gain in body weight and in tibia epiphyseal width was observed in animals inoculated with rcGH-pN. Thus it is concluded that recombinant plasmid of cGH cDNA may be used as supplement to increase the meat production. The effects of the same plasmid on milk production may be checked in future experiments using Beetal goat. Copyright 2014 Friends Science Publishers