ACTA VETERINARIA ET ZOOTECHNICA SINICA ›› 2010, Vol. 41 ›› Issue (10): 1232-1239.doi:

• 遗传繁育 • Previous Articles     Next Articles

Identification and Prokaryotic Expression of Duck Adiponectin Gene

XUE Mao-yun2, DONG Biao3, ZHANG Ying1, YU Jian-feng1, MENG He4, GONG Dao-qing5,   

  1. 1. Department of Biology, Changshu Institute of Technology, Changshu 215500, China; 2. Jiangsu Institute of Economic and Trade Technology, Nanjing 211168,China; 3. Jiangsu Animal Husbandry Veterinary College, Taizhou 225300, China; 4. College of Agriculture and Biology, Shanghai Jiao Tong University, Shanghai 200240, China; 5. College of Animal Science and Technology, Yangzhou University, Yangzhou 225009, China
  • Received:1900-01-01 Revised:1900-01-01 Online:2010-10-25 Published:2010-10-25

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Abstract: The purpose of this study was to clone and characterize the duck Adiponectin gene, and to reveal the tissue expression pattern of Adiponectin mRNA in duck, and to express recombinant duck Adiponectin in prokaryotic cells. The sequence of duck Adiponectin gene was obtained by reverse transcription PCR, 5 ′-rapid amplify cDNA end and 3′- rapid amplify cDNA end (5′-RACE and 3′-RACE) methods, and its expression in various duck tissues was detected by semi-quantity RT-PCR. Recombinant plasmid was constructed and transformed into E.coli. to express the recombinant duck Adiponectin in prokaryotic cells. The three Adiponectin cDNA fragments were amplified from total RNA using RT-PCR, 5′-RACE and 3′-RACE methods, respectively. After cloning and sequencing the three fragments, a 1 374 bp full length cDNA sequence of duck Adiponectin gene was acquired, which contained a full 738 bp nucleotides open reading frame, encoding 245 amino acids. The deduced Adiponectin amino acid sequence of duck contained 22 glycine-X-Y repeats at the collagen repeats as found in the chicken and mammalian Adiponectin. The duck Adiponectin CDS shared 94.9% and 86.0% homology with goose and chicken, and about 64.2%-67.0% homology with mouse, human and dog. Meanwhile, the putative amino acid sequence of duck Adiponectin shares 95.5% and 84.0% homology with goose and chicken, and about 65% homology with the above mammals. The semi-quantity RT-PCR showed that Adiponectin expressions were detectable in all tissues, and the expression level of Adiponectin gene was high in adipose, muscular stomach, small intestine, heart and skeletal muscle, medium in lung, gland stomach, ovary and spleen, low in liver, kidney and diencephalons. The recombinant plasmid of pET41a-duck-adp was constructed and transformed into E. coli BL21（DE3）. The 30 ku recombinant duck adiponectin was successfully expressed in E. coli. The Adiponectin gene is conserved in animal evolution, and Adiponectin mRNA expresses differently in various tissues. The recombined duck adiponectin could be expressed in prokaryotic system.