ACTA VETERINARIA ET ZOOTECHNICA SINICA ›› 2009, Vol. 40 ›› Issue (5): 611-616.doi:

• 遗传繁育 •     Next Articles

Cloning of Porcine Suv39h2 cDNA and Its Expression in E. coli

LIU Li-na, PENG Jian, ZHENG Rong, LIU Min, JIANG Si-wen*   

  1. Key Laboratory of Swine Genetics and Breeding of Ministry of Agriculture & Key Laboratory of Agricultural Animal Genetics, Breeding, and Reproduction of Ministry of Education, Huazhong Agricultural University, Wuhan 430070,China
  • Received:1900-01-01 Revised:1900-01-01 Online:2009-05-25 Published:2009-05-25

Abstract: Based on the mRNA sequence of human Suv39h2 gene, a porcine fragment of its cDNA sequence was obtained using in silico cloning method. By RT-PCR method, a porcine Suv39h2 cDNA fragment of 1 291 bp was amplified from muscle tissue and inserted into pGEX-KG plasmid expressing glutathione Stransferase (GST) fusion protein. After confirmation by the sequencing and restriction enzyme analysis, the recombinant plasimids were transformed into E. coli BL21 competent cells. The optimal IPTG induction conditions were determined as follows: induction time was 4 h and IPTG concentration was 0.7 mmol·L-1. The fusion protein was insoluble. Using Western blotting, we found a band with molecular weight of 66 kD, which indicated that porcine Suv39h2 was expressed in E. coli. This work laid a foundation for further work on functional study.