Cloning of human chorionic gonadotropin beta subunit in gene transfer via sperm rooster

Background: The poultry production technology in animal transgenic has taken into consideration for having many advantages such as short generation times, the large number of production of offspring and suitable pattern of protein glycosylation. HCG hormone is one of the most important hormones that can be produced into recombinant in transgenic poultry. This hormone composed of two subunits, alpha and beta. The beta subunit is responsible for the specific activity of this hormone and it makes it specific. The aim of this study was cloning of the beta subunit of human chorionic gonadotropin. It was in an appropriate vector through sperm that could be used in the production of transgenic poultry.Methods: For this purpose, the hormone beta subunits were amplified by specific primer pairs, and were cloned into T vector in restriction sites XhoI and HindIII. The recombinant plasmid was transformed into competent E. coli cells and colonies that containing recombinant plasmids were selected by colony PCR. The validity of extracted plasmid of the clones was analyzed by enzyme digestion and sequencing. The beta chain of T vector was isolated and was cloned again into pcDNA3.1 + expression vector.Result: The results of enzyme analysis and sequencing indicated that recombinant plasmid pCDNA3.1 +/βhCG was cloned with the correct sequence and completely matched up with human chorionic gonadotropin beta sub unit gene.Conclusion: The cloned gene due to having the correct sequence was used in gene transfering process via rooster sperm.

Keywords:

Plasmid , Sequencing , Sperm , Transgenic chickens , Cloning