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HEREDITAS ›› 2008, Vol. 30 ›› Issue (11): 1433-1438.doi: 10.3724/SP.J.1005.2008.01433

• 研究报告 • Previous Articles     Next Articles

Cloning and optimizing prokaryotic induced expression conditions of prolactin in White Goose

GUO Li1;YANG Huan-Min1;LI Peng1;KANG Bo2   

  1. 1. College of Animal Science and Veterinary Medicine, Heilongjiang August First Land Reclamation University, Daqing 163319, China;
    2. College of Animal Science and Veterinary Medicine, Jilin University, Changchun 130062, China
  • Received:2008-01-20 Revised:2008-04-19 Online:2008-11-10 Published:2008-11-10
  • Contact: YANG Huan-Min

Abstract: Abstract: The mature segment gene of prolactin (PRL) in White Goose was amplified from pituitary by RT-PCR and then cloned into the pMD18-T vector. Sequencing analysis showed that the cDNA has a length of 690 bp including the termina-tion codon and encodes a protein composed of 230 amino acids, which differs from the published PRL cDNA sequence. There is a homology of 99.57% in base and 99.56% in amino acids with that of Wanxi White Goose, respectively. A pro-karyotic expression vector, pET-32a(+), was used to construct the recombinant plasmid pET-32a(+)-PRL to produce protein. Having been induced by IPTG,the host cell carrying the recombinant plasmid expressed the recombinant PRL. The opti-mal condition for expression is 1 mmol/L IPTG at 37℃. Based on this condition, the expression rose to the highest level by 4 h of induction, accounting for 28.96% of the total bacterial protein.