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Cloning, expression and tissue distribution of IGF-1 from giant panda (Ailuropoda melanoleuca)

Published online by Cambridge University Press:  09 March 2007

X. -L. Hu
Affiliation:
College of Life Sciences, Zhejiang University, Hangzhou 310058, China Geriatrics Institute, Zhejiang Hospital, Hangzhou 310013, China
M. -Y. Zhu*
Affiliation:
College of Life Sciences, Zhejiang University, Hangzhou 310058, China
Z. -H. Zhang
Affiliation:
Key Laboratory for Reproduction and Conservation Genetics of Endangered Wildlife of Sichuan Province, Chengdu Research Base of Giant Panda Breeding, Chengdu 610081, China
R. Hou
Affiliation:
Key Laboratory for Reproduction and Conservation Genetics of Endangered Wildlife of Sichuan Province, Chengdu Research Base of Giant Panda Breeding, Chengdu 610081, China
F. -J. Shen
Affiliation:
Key Laboratory for Reproduction and Conservation Genetics of Endangered Wildlife of Sichuan Province, Chengdu Research Base of Giant Panda Breeding, Chengdu 610081, China
A. -J. Zhang
Affiliation:
Key Laboratory for Reproduction and Conservation Genetics of Endangered Wildlife of Sichuan Province, Chengdu Research Base of Giant Panda Breeding, Chengdu 610081, China
*
Corresponding author: E-mail: [email protected]
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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.

Type
Research Article
Copyright
Copyright © British Society of Animal Science 2006

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