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Cloning of the obese gene (ob) of adipose tissue, and differences in ob gene expression with age of piglets

Published online by Cambridge University Press:  12 February 2007

Wang Yi-Zhen ,
Chu Xiao-Na ,
Huang Hai-Qing ,
Han Fei-Fei  and
Liu Jian-Xin
Wang Yi-Zhen*
Affiliation:
Institute of Feed Science, Zhejiang University, Hangzhou 310029, China
Chu Xiao-Na
Affiliation:
Institute of Feed Science, Zhejiang University, Hangzhou 310029, China The Endocrinology and Diabetes Hospital of Qingdao, Qingdao 266071, China
Huang Hai-Qing
Affiliation:
Institute of Feed Science, Zhejiang University, Hangzhou 310029, China
Han Fei-Fei
Affiliation:
Institute of Feed Science, Zhejiang University, Hangzhou 310029, China
Liu Jian-Xin
Affiliation:
College of Animal Science, Zhejiang University, Hangzhou 310029, China
*
*Corresponding author: Email: yzwang@zju.edu.cn
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Abstract

Genomic RNA was extracted from the subcutaneous adipose tissue of piglets (Duroc×Landrace×Tai-hu) at 1, 14, 28, 42 and 56 days of age, and obese gene (ob) mRNA was amplified using reverse transcriptase-polymerase chain reaction (RT-PCR). A DNA fragment of about 504 bp was obtained and the PCR product was cloned into a pGEM-T vector. The ob gene was isolated and sequenced from the positive clones screened. Sequence analysis suggested that this fragment was a partial sequence of ob cDNA, coding 167 amino acid residues, which constituted the major part of leptin mature protein. The gene homology of the fragment obtained in this study compared to the reported ob cDNA sequence in adipocytes of pig was 99.405%, and amino acid homology was 98.94%. Based on the ob gene clone, we successfully constructed an optimal semi-quantitative RT-PCR method. Using β-actin as the internal control, we investigated the difference of ob gene expression at different ages of piglets. Results showed that ob mRNA levels increased steadily at postnatal days 1–28 (preweaning), peaked at postnatal day 28, when piglets were weaned, and decreased from day 28 to 56.

Keywords

clonegene expressionobese genepigletsRT-PCR
Type
Research Article
Information
Chinese Journal of Agricultural Biotechnology , Volume 2 , Issue 3 , December 2005 , pp. 207 - 212
Copyright
Copyright © China Agricultural University and Cambridge University Press 2005

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References

Barb, CR, Hausman, GJ and Houseknecht, KL (2001) Biology of leptin in the pig. Domestic Animal Endocrinology 21: 297–317.CrossRefGoogle ScholarPubMed
Bidwell, CA, Ji, SQ, Frank, GR and Cornelius, SG (1997) Cloning and expression of the porcine obese gene. Animal Biotechnology 8(2): 191–206.Google Scholar
Chen, D and Klebe, RJ (1993) Controls for validation of relative reverse transcription–polymerase chain reaction assays. PCR Methods Application 3: 127–129.CrossRefGoogle ScholarPubMed
Debao, L and Ping, X (1994) The Principles and Methods of Recombinant DNA. Hangzhou: Zhejiang Science and Technology Press (in Chinese).Google Scholar
Frederich, MA (1998) Short Protocols in Molecular Biology. Beijing: Science Press (in Chinese).Google Scholar
Li, H, Matheny, M, Tumer, N and Scarpace, PJ (1998) Aging and fasting regulation of leptin and hypothalamic neurpeptide Y gene expression. American Journal of Physiology 275: E405–E411.Google ScholarPubMed
Lion, T (1996) Control genes in reverse transcriptase polymerase chain reaction assays. Leukemia 10: 1527–1528.Google Scholar
Marone, M, Mozzetti, S, Ritis, DD, Pierelli, L and Scambia, G (2001) Semiquantitative RT-PCR analysis to assess the expression levels of multiple transcripts from the same sample. Biological Proceedings Online 3(1): 19–25.CrossRefGoogle ScholarPubMed
Nogalska, A and Swierczynski, JL (2001) The age-related differences in obese and fatty acid synthase gene expression in white adipose tissue of rat. Biochimica et Biophysica Acta 1533: 73–80.Google Scholar
Ramsay, TG, Yan, X and Morrison, C (1998) The obesity gene in swine: sequence and expression of porcine leptin. Journal of Animal Science 76: 484–490.CrossRefGoogle ScholarPubMed
Rayner, DV, Dalgliesh, GD, Duncan, JS, Hardie, LJ, Hoggard, N and Trayhurn, P (1997) Postnatal development of the ob gene system: Elevated leptin levels in suckling fa/fa rats. American Journal of Physiology 273: R446–R450.Google Scholar
Rentsch, J and Chiesi, M (1996) Regulation of ob gene mRNA levels in cultured adipocytes. FEBs Letters 379: 55–59.Google Scholar
Robert, VC and Jose, FC (1997) Leptin and the regulation of body weight. International Journal of Biochemistry and Cell Biology 29(11): 1255–1272.Google Scholar
Spurlock, ME, Frank, GR and Cornelius, SG (1998) Obese gene expression in porcine adipose tissue is reduced by food deprivation but not by maintenance or submaintenance food intake. Journal of Nutrition 128: 677–682.CrossRefGoogle Scholar
Wang, BL, Shao, JL and Zheng, JN (1989) The research report on the growth and development rule in commercial leaning pigs. Ningxia Journal of Agriculture and Forest Technology 5: 34–38.Google Scholar