Journal of Animal Production, Print ISSN: 2008-6776., Online ISSN: 2382-994X

Cloning of human chorionic gonadotropin beta subunit for rooster sperm mediated gene transfer

Document Type : Research Paper

Authors

Mehdi Noorani 1
Shaban Rahimi 2
Abdolhoseein Shahverdi 3
Mohsen Sharafi 4

1 MSc in Poultry Science, Tarbiat Modares University

2 Professor Department of Poultry Science, Faculty of Agriculture, Tarbiat Modares University

3 Faculty member at Royan Institute

4 Faculty member at the Department of Poultry Science, Tarbiat Modares University

https://doi.org/10.22059/jap.2018.208607.623055
Abstract
The aim of this study was cloning of the beta subunit of human chorionic gonadotropin in an appropriate vector for production of transgenic chicken trrough sperm mediated gene transfer. In this regard, transgenic chicken production tecnology 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. To date, no study has been conducted on the cloning of the beta subunit of human chorionic gonadotropin for rooster sperm. For this purpose, the hormone beta subunit were amplified by a specific primer pairs, and cloned in T vector. 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 were analyzed by enzyme digestion and sequencing. The beta chain of T vector was isolated and was cloned again into pcDNA3.1 + expression vector. The results of enzyme analysis and sequencing indicated that recombinant plasmid pCDNA3.1 +/βhCG were cloned with the correct sequence and completely matched up with human chorionic gonadotropin beta subunit gene that can be concluded that it has sutible stracture for sperm mediated gene transfer.

Keywords

plasmid
Sperm
sequencing
transgenic chicken
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Journal of Animal Production, Print ISSN: 2008-6776., Online ISSN: 2382-994X
Volume 20, Issue 2
Summer 2018
Pages 203-212

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