ACTA VETERINARIA ET ZOOTECHNICA SINICA ›› 2007, Vol. 38 ›› Issue (7): 625-629.doi:

• 遗传繁育 •     Next Articles

Molecular Cloning, Sequence Analysis and Prokaryotic Expression of the Porcine Sar1b Gene

WANG Xue-min；LIU Bang；ZHAO Shu-hong；FAN Bin；ZHU Meng-jin；YU Mei；XIONG Tong-an；LI Kui   

  1. 1.Key Laboratory of Animal Genetics, Breeding and Reproduction of Ministry of Education,Huazhong Agricultural University, Wuhan 430070, China；2.Institute of Animal Science, Jiangsu Academy of Agricultural Sciences, Nanjing 210014, China；3.Institute of Animal Science, Chinese Academy of Agricultural Sciences, Beijing 100094, China
  • Received:1900-01-01 Revised:1900-01-01 Online:2007-07-25 Published:2007-07-25

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Abstract: A pair of primers was designed based on the published nudeotide sequence of the porcine Sar1b gene (GenBank accession number: AY819557). Total RNA was isolated from the pig liver tissue and the first-strand cDNA was generated using reverse transcription reactions. The full-length coding sequence of the porcine Sar1b cDNA was amplified by RT-PCR. The product was double-digested by EcoRI and SalI, and then directionally cloned into the pET28a vector. The recombinant expression plasmid in the E. coli BL21 (DE3) was induced with 1 mmol/L ITPG. SDS-PAGE analysis showed that the induced expressed protein was about 26 ku. Western blot revealed that the expressed protein was the His tag fusion protein, which indicated that the recombinant prokaryotic expression vector expressed the fusion protein successfully. The cloning and expression of the porcine Sar1b gene established the foundation for further study on its biological function.