ACTA VETERINARIA ET ZOOTECHNICA SINICA ›› 2009, Vol. 40 ›› Issue (12): 1712-1717.doi:

• 遗传繁育 • Previous Articles     Next Articles

CIB1 cDNA Cloning from Small Tail Han Sheep and Its Expression in E.coli

YU Yan 1, 2, WANG Chuduan 1, LI Hongbin 3, WEI Caihong 3, SUN Dan 4, LU Jian 5, LIU Kaidong 6, LV Yanfei 6, DU Lixin 3*
  

  1. 1. College of Animal Science and Technology, China Agricultural University, Beijing 100193, China; 2. Reproductive Medical Center, The Second Hospital Affiliated to Shandong University of Traditional Chinese Medicine, Jinan 250001, China; 3. Institute of Animal Science, Chinese Academy of Agricultural Sciences, Beijing 100193, China; 4. College of Life Sciences, Liaoning Normal University, Dalian 116029, China; 5. College of Animal Science and Technology, Yangzhou University,Yangzhou 225009, China; 6. College of Animal Science＆Veterinary Medicine, Qingdao Agricultural University, Qingdao 266109, China
  • Received:2008-12-22 Revised:1900-01-01 Online:2009-12-24 Published:2009-12-24
  • Contact: DU Lixin

PDF (PC)

608

Abstract: In order to clone cDNA of Small Tail Han sheep CIB1 gene, and express the fusion protein in E.coli, a pair of specific primers were designed according to the nucleotide sequence of the published CIB1 gene. Fulllength cDNA of CIB1 gene was cloned from Small Tail Han sheep by RTPCR. CIB1 was cloned into pMD19T vector and identified by sequencing. CIB1 gene was cloned into pET32a vector to construct a prokaryotic expression vector pET32aCIB1 and transformed into BL21（DE3）pLysS cells. The recombinant plasmid pET32aCIB1 was induced in Escherichia coli using IPTG. The results showed that the identity at nucleotide level were more than 90% with CIB1 gene from cattle, pig, rhesus monkey, human, mouse, rat, chimpanzee and so on, and the amino acid similarity were higher than 93% correspondingly. The CIB1 cDNA containing a 576 bp open reading frame (ORF) encoding 191 amino acids. The expressed recombinant fusion protein is 38 ku in size. These results provided basis for the future research on ovine CIB1 protein function.