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蓝塘猪和大白猪消减cDNA文库与差异基因分析
引用本文:蔡更元,陈瑶生,李加琪,张豪,王翀.蓝塘猪和大白猪消减cDNA文库与差异基因分析[J].中国农业科学,2011,44(12):2553-2560.
作者姓名:蔡更元  陈瑶生  李加琪  张豪  王翀
作者单位:华南农业大学;
基金项目:广东省自然科学基金重点项目(9251064201000005); 广东省科技攻关项目(2009B020306002); 广州市科技支撑计划(07-支撑-05)
摘    要: 【目的】寻找肉质性状候选优势表达基因,研究猪肉质优良性状的遗传基础和分子机制。【方法】利用抑制性消减杂交技术构建蓝塘猪和大白猪肌肉组织差异表达的消减cDNA文库,利用实时荧光定量PCR法研究基因的差异表达。【结果】获得61个有效克隆。对整个文库进行测序分析,获得47条有效序列。用BLAST在线软件与GenBank 和GenBank EST等数据库进行同源序列比较,发现其中有2条未知序列。对文库中所包含的MYL1、LDHA、KPNA3、TPM3、HUMMLC2B、GNTN和Oaz等基因进行了实时荧光定量PCR检测,结果显示MYL1、LDHA、KPNA3、TPM3和HUMMLC2B这5个基因在蓝塘猪肌肉组织的表达量显著高于大白猪(P<0.05);GNTN和Oaz基因在大白猪肌肉组织中的表达量显著高于蓝塘猪(P<0.05)。【结论】很多EST与肉质有关的重要基因具有很高的同源性,这些基因在蓝塘猪和大白猪中表达差异显著(P<0.05)。

关 键 词:SSH  肌肉组织  差异表达基因  消减cDNA文库  
收稿时间:2010-07-03;

Isolation of Differentially Expressed Genes between Lantang Pig and Large White Pig by SSH and Q-PCR
CAI Geng-yuan,CHEN Yao-sheng,LI Jia-qi,ZHANG Hao,WANG Chong.Isolation of Differentially Expressed Genes between Lantang Pig and Large White Pig by SSH and Q-PCR[J].Scientia Agricultura Sinica,2011,44(12):2553-2560.
Authors:CAI Geng-yuan  CHEN Yao-sheng  LI Jia-qi  ZHANG Hao  WANG Chong
Institution:CAI Geng-yuan1,2,CHEN Yao-sheng1,3,LI Jia-qi1,ZHANG Hao1,WANG Chong1(1South China Agricultural University,Guangzhou 510640,2Institute of Animal Science,Guangdong Academy of Agricultural Sciences,Guangzhou 510642,3Sun Yat-sen University,Guangzhou 510006)
Abstract:【Objective】The aim of this experiment is to investigate the genes related to meat quality, and study its genetic basis and molecular mechanism.【Method】A subtracted cDNA library was constructed by using suppression subtractive hybridization, differential expressions of the genes were studied by using real-time quantitative PCR. 【Result】A total of 61 positive clones were screened. A total of 47 effective sequences were found by sequencing in the whole subtractive cDNA library. After comparisons with GenBank, GenBank ESTs librarys using the online software of the BLAST, the two unknown sequences were found. The expressions of the MYL1, LDHA, KPNA3, TPM3, HUMMLC2B, GNTN, and Oaz genes were studied by real-time quantitative PCR. The results showed that significant differences were found between the Lantang Pig and Large White Pig. The expressions of the MYL1, LDHA, KPNA3, TPM3, and HUMMLC2B genes were significantly higher in the Lantang Pig (P<0.05), and the expressions of the GNTN and Oaz genes were significantly higher in the Large White Pig (P<0.05). 【Conclusion】Many of these ESTs were highly homologous with the important genes related to meat quality traits, and significant differences of them were found between Lantang Pig and Large White Pig.
Keywords:SSH  muscle tissue  differentially expressioned gene  subtractive cDNA library  
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