Animal Science Abstract
Cloning, expression and tissue distribution of IGF-1 from giant panda (Ailuropoda melanoleuca)
X.-L. Hu1,2, M.-Y. Zhu1,A, Z.-H. Zhang3, R. Hou3, F.-J. Shen3 and A.-J. Zhang3
ACorresponding author. E-mail: myzhu@zju.edu.cn
1College of Life Sciences, Zhejiang University, Hangzhou 310058, China2Geriatrics Institute, Zhejiang Hospital, Hangzhou 310013, China
3Key Laboratory for Reproduction and Conservation Genetics of Endangered Wildlife of Sichuan Province, Chengdu Research Base of Giant Panda Breeding, Chengdu 610081, China
Abstract
Insulin-like growth factor-1 (IGF-1) is a polypeptide that regulate growth, differentiation and muscle regeneration in several species. This study was designed to analyse the cDNA sequence of IGF-1 in giant panda, its gene expression and tissue distributions in relation to growth, development and reproduction of captive giant pandas. A cDNA encoding IGF-1 was cloned by reverse transcriptase-PCR from a male giant panda cub liver tissue. Sequence alignment analysis showed the cysteine residues and other amino acid residues of the mature IGF-1 peptide of giant panda are highly conserved in mammalian species. DNA encoding the mature IGF-1 peptide of the giant panda was ligated with pET-DsbA expression vector and expressed in E. coli BL21. Western blot confirmed the IGF-1 antigen activity of the recombinant protein. By in situ hybridization and immunohistochemistry analysis it is demonstrated that the IGF-1 mRNA distribution abundance were significantly higher in the liver, muscles and heart than those in the other tissues studied. However, the IGF-1 peptide distribution abundances in all the tissues studied did not show any significant differences. The localization of the IGF-1 peptide was in accordance with that of mRNA for each tissue. These results may suggest that IGF-1 play an autocrine or paracrine rôle in regulation of the giant panda growth and development.
Keywords: Ailuropoda melanoleuca, cDNA cloning, gene expression, insulin-like growth factor-1, tissue distribution.
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