Scientia Agricultura Sinica ›› 2012, Vol. 45 ›› Issue (15): 3154-3164.doi: 10.3864/j.issn.0578-1752.2012.15.018

• ANIMAL SCIENCE·RESOURCE INSECT • Previous Articles     Next Articles

Construction of Subtractive cDNA Library of Skin Tissue in Follicle Anagen Between Chinese Merino and Kazakh Sheep

 YANG  Jian-Bo, GAN  Shang-Quan, LI  Jing, GAO  Lei, YANG  Jing-Quan, ZHANG  Ke, YANG  Yong-Lin, SHEN  Min, SHI  Guo-Qing   

  1. 1.石河子大学动物科技学院，新疆石河子 832000
    2.新疆农垦科学院畜牧兽医研究所，新疆石河子 832000
    3.新疆兵团绵羊繁育生物技术重点实验室，新疆石河子 832000
  • Received:2012-02-20 Online:2012-08-01 Published:2012-05-03

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Abstract: 【Objective】The aim of the study is to screen important candidate genes influencing wool traits in sheep, and to lay a theoretical foundation for exploring the molecular mechanism of wool traits formation.【Method】Using suppression subtractive hybridzation (SSH) technique, positive and reverse subtracted cDNA library between skin tissue in follicle anagen of Chinese Merino (superfine type) and Kazakh sheep was constructed, respectively. Genes of differential expression profiles were sequenced for bioinformatics analysis. Semi-quantitative RT-PCR and real-time fluorescent quantitative RT-PCR methods were used to assess partial genes of differential expression profiles.【Result】 A total of 153 and 143 ESTs were obtained from the forward and reverse subtracted cDNA library, respectively. Among these ESTs, 5 from positive library and 4 from reverse library of unknown function were identified and speculated as novel genes. The GO cluster, pathway and protein-protein interaction analysis of the partial ESTs of known function showed that there were certain differences between coarse wool and fine wool sheep. The three differential genes from the constrcted cDNA libraries, KRTAP8-2, Trichohyalin and KRTAP3-2 genes were all specifically expressed at high level in the skin. The abundance of KRTAP8-2 and Trichohyalin genes mRNA in skin of Chinese Merino was 3.45 and 2.07 times than that of Kazakh sheep, respectively, while the abundance of KRTAP3-2 gene mRNA in skin of Kazakh sheep was 1.87 times than that of Chinese Merino. 【Conclusion】The subtracted cDNA library of skin tissue between Chinese Merino and Kazakh sheep was successfully constructed, a batch of ESTs with differential expression profiles which possibly influence wool traits were initially screened.

Key words: suppression subtractive hybridzation, Chinese Merino sheep, Kazakh sheep, skin

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