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Isolation, cloning and sequence analysis of differentially expressed genes in the backfat tissue between hybrids and parents in pigs

Published online by Cambridge University Press:  12 February 2007

Ren Zhu-Qing ,
Xiong Yuan-Zhu ,
Deng Chang-Yang ,
Lei Ming-Gang ,
Zuo Bo ,
Zheng Rong ,
Li Feng-E ,
Jiang Si-Wen  and
Jiang Xun-Ping
Ren Zhu-Qing
Affiliation:
Key Laboratory of Swine Breeding and Genetics, Ministry of Agriculture, Huazhong Agricultural University, Wuhan 430070, China
Xiong Yuan-Zhu*
Affiliation:
Key Laboratory of Swine Breeding and Genetics, Ministry of Agriculture, Huazhong Agricultural University, Wuhan 430070, China
Deng Chang-Yang
Affiliation:
Key Laboratory of Swine Breeding and Genetics, Ministry of Agriculture, Huazhong Agricultural University, Wuhan 430070, China
Lei Ming-Gang
Affiliation:
Key Laboratory of Swine Breeding and Genetics, Ministry of Agriculture, Huazhong Agricultural University, Wuhan 430070, China
Zuo Bo
Affiliation:
Key Laboratory of Swine Breeding and Genetics, Ministry of Agriculture, Huazhong Agricultural University, Wuhan 430070, China
Zheng Rong
Affiliation:
Key Laboratory of Swine Breeding and Genetics, Ministry of Agriculture, Huazhong Agricultural University, Wuhan 430070, China
Li Feng-E
Affiliation:
Key Laboratory of Swine Breeding and Genetics, Ministry of Agriculture, Huazhong Agricultural University, Wuhan 430070, China
Jiang Si-Wen
Affiliation:
Key Laboratory of Swine Breeding and Genetics, Ministry of Agriculture, Huazhong Agricultural University, Wuhan 430070, China
Jiang Xun-Ping
Affiliation:
Key Laboratory of Swine Breeding and Genetics, Ministry of Agriculture, Huazhong Agricultural University, Wuhan 430070, China
*
*Corresponding author: E-mail: [email protected]
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Abstract

In order to reveal the molecular basis of heterosis, Large White (LW), an introduced European pig breed, and Meishan (MS), a Chinese indigenous pig breed, were selected to hybridize directly and reciprocally in the present experiment. mRNA differential display (DD) technique was performed to identify genes that were differentially expressed in the backfat tissues of hybrids (LW×MS, MS×LW) and purebred (LW×LW, MS×MS) pigs. The ten anchor primers in combination with ten arbitrary primers (100 sets in total) were used and nearly 1500 reproducible bands were observed in polyacrylamide gels. The 40 differentially displayed bands were selected for cloning and sequencing. Thirty-six out of 40 expressed sequence tags (ESTs) proved to be novel and the sequences were submitted to GenBank (accession No. CV507051-CV507087); the other four showed similarity to known genes published in GenBank. Three among 36 novel ESTs were chosen for further identification with semi-quantitative reverse transcriptase polymerase chain reaction (RT-PCR). The result showed that two ESTs were differentially expressed, and the third showed no obvious difference between hybrids and purebreds. In order to reduce the percentage of false-positive DD, RNA pools of four types of pigs were constructed, by mixing samples from six pigs of the same genotype, and subjected to DD. Stringent annealing temperature was applied and only bands that could be repeated in duplicate PCR were used for further study. The results showed that the expression pattern of these 36 ESTs differed among the four genotypes of pigs, suggesting that the genes corresponding to these differentially expressed ESTs might be related to the heterosis occurring in fat tissue.

Keywords

ESTsheterosismRNA differential displaypigsemi-quantitative RT-PCR
Type
Research Article
Information
Chinese Journal of Agricultural Biotechnology , Volume 3 , Issue 1 , April 2006 , pp. 47 - 52
Copyright
Copyright © China Agricultural University and Cambridge University Press 2006

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References

Conover, CA (1991) A unique receptor-independent mechanism by which insulin-like growth factor I regulates the availability of insulin-like growth factor binding proteins in normal and transformed human fibroblasts. Journal of Clinical Investigation 88: 1354–1361.CrossRefGoogle ScholarPubMed
Dahle, MK, Gronning, LM, Cederberg, A et al. , (2002) Mechanisms of FOXC2- and FOXD1-mediated regulation of the RI alpha subunit of cAMP-dependent protein kinase include release of transcriptional repression and activation by protein kinase B alpha and cAMP. Journal of Biological Chemistry 277: 22902–22908.CrossRefGoogle ScholarPubMed
Davoli, R, Zambonelli, P, Bigi, D, Fontanesi, L and Russo, V (1990) Analysis of expressed sequence tags of porcine skeletal muscle. Gene 233: 181–188.CrossRefGoogle Scholar
Johns, GF (1998) Genetic aspacts of domestication, common breeds and their origin. In: Rothchild, MF and Ruvinsky, A (editors) The Genetics of the Pig. Wallingford, UK: CAB International, pp. 17–48.Google Scholar
Kern, PA, Saghizadeh, M, Ong, JM, Bosch, RJ, Deem, R and Simsolo, RB (1995) The expression of tumor necrosis factor in human adipose tissue. Regulation by obesity, weight loss, and relationship to lipoprotein lipase. Journal of Clinical Investigation 95: 2111–2119.CrossRefGoogle ScholarPubMed
Liang, P and Pardee, AB (1992) Differential display of eukaryotic messenger RNA by means of the polymerase chain reaction. Science 257: 967–970.CrossRefGoogle ScholarPubMed
Liang, P, Averboukh, L and Pardee, AB (1993) Distribution and cloning of enkaryotic mRNA by means of differential display: Refinements and optimization. Nucleic Acids Research 21: 3269–3275.CrossRefGoogle Scholar
Liang, P, Zhu, W, Zhang, X et al. , (1994) Differential display using one-base anchored oligo-dT primers. Nucleic Acids Research 22: 5763–5764.CrossRefGoogle ScholarPubMed
Liu, GY, Xiong, YZ, Deng, CY, Zuo, B and Zhang, JH (2004) Comparison of gene expression patterns in longissimus dorsi of pigs between the High-parent heterosis cross combination landrace Large White and the mid-parent heterosis cross combination Large White–Meishan. Asian-Australian Journal of Animal Science 17: 1192–1196.CrossRefGoogle Scholar
Luheshi, GN, Gardner, JD, Rushforth, DA, Loudon, AS and Rothwell, NJ (1999) Leptin actions on food intake and body temperature are mediated by IL-1. Proceedings of the National Academy of Sciences, USA 96: 7047–7052.CrossRefGoogle ScholarPubMed
Meadus, WJ (2003) A semi-quantitative RT-PCR method to measure the in vivo effect of dietary conjugated linoleic acid on porcine muscle PPAR gene expression. Biological Procedures Online 5: 20–28.CrossRefGoogle ScholarPubMed
Romagnoli, S, Maddaloni, M, Livini, C and Motto, M (1990) Relationship between gene expression and hybrid vigor in primary root tips of young maize (Zea mays L.) plantlets. Theory of Applied Genetics 80: 767–775.CrossRefGoogle ScholarPubMed
Tsaftaris, AS (1995) Molecular aspects of heterosis in plants. Physiologia Plantarum 94: 362–370.CrossRefGoogle Scholar
Wang, D, Zhang, Y, Sun, DX, Yu, Y, Xu, GY and Li, JY (2004a) Gene differential expression of liver tissues in crossbred versus purebred chicken and their relationship with heterosis of meat trait. Acta Genetica Sinica 31: 257–264.Google ScholarPubMed
Wang, YZ, Lin, WX and Xu, ZR (2004b) Antibacterial peptide pr-39 gene expression in different growth stages of pig evaluation by semi-quantitative RT-PCR method. Journal of Agricultural Biotechnology 12: 52–55.Google Scholar
Wu, LM, Ni, ZF and Sun, QX (2001) Relationship between differential expression patterns of multigene families and heterosis in a wheat diallel cross. Acta Genetica Sinica 28: 256–266.Google Scholar
Xie, XD, Ni, ZF, Meng, FR, Wu, LM, Wang, ZK and Sun, QX (2003) Relationship between difference of gene expression in early developing seeds of hybrid versus parents and heterosis in wheat. Acta Genetica Sinica 30: 260–266.Google ScholarPubMed
Xiong, LZ, Yang, GP and Zhang, QF (1998) Relationship of differential gene expression in leaves with heterosis and heterozygosity in a rice diallel cross. Molecular Breeding 4: 129–136.CrossRefGoogle Scholar
Yamamoto, K, Takeshita, K, Shimokawa, T et al. , (2002) Lasminogen activator inhibitor-1 is a major stress-regulated gene: Implications for stress-induced thrombosis in aged individuals. Proceedings of the National Academy of Sciences, USA 99: 890–895.CrossRefGoogle Scholar
Yamauchi, T (2001) The fat-derived hormone adiponectin reverses insulin resistance associated with both lipoatrophy and obesity. Natural Medicine 5: 20–28.Google Scholar
Yumoto Hiromichi, , Nakae Hideaki, , Fujinaka Keiko, , Ebisu Shigeyuki, and Matsuo Takashi, (1999) Interleukin-6 (IL-6) and IL-8 are induced in human oral epithelial cells in response to exposure to periodontopathic Eikenella corrodens. Infection and Immunity 67(1): 384–394.CrossRefGoogle Scholar