作　　者： ; ; ; ; ; ;

机构地区： 华中农业大学动物科学技术学院农业部猪遗传育种重点开放实验室

出　　处： 《Acta Genetica Sinica》 2003年第7期668-672,共5页

摘　　要： 以梅山猪和长白猪的背最长肌为材料 ,利用抑制性消减杂交技术成功构建了梅山猪与长白猪肌肉组织间正反向消减cDNA文库。以管家基因G3PDH作为消减指标 ,检测梅山猪和长白猪两个消减文库的消减效率分别高达 2 10 和 2 5倍 ,表明某些特异于两种肌肉组织的差异表达基因的富集效率也分别接近 2 10 和 2 5倍。从两个消减文库中分别筛选到了 70 9和 6 73个有效阳性克隆 ,PCR鉴定插入片段长度主要分布于 15 0～ 75 0bp之间。不同猪种肌肉组织间消减cDNA文库的构建为进一步分离和鉴定与猪肌肉生长和肉质相关基因奠定了基础 ,对探讨肌肉生长机理和肉质决定因素以及猪的分子育种具有重要意义。 Using suppression subtractive hybridization (SSH) technique,forward and reverse subtracted cDNA libraries were constructed between Longissimus muscles from Meishan and Landrace pigs.A housekeeping gene, G3PDH ,was used to estimate the efficiency of subtractive cDNA.In two cDNA libraries, G3PDH was subtracted very efficiently at appropriate 2 10 and 2 5 folds,respectively,indicating that some differentially expressed genes were also enriched at the same folds and the two subtractive cDNA libraries were very successful.A total of 709 and 673 positive clones were isolated from forward and reverse subtracted cDNA libraries,respectively.Analysis of PCR showed that most of all plasmids in the clones contained 150～750 bp inserts.The construction of subtractive cDNA libraries between muscle tissue from different pig breeds laid solid foundations for isolating and identifying the genes determining muscle growth and meat quality,which will be important to understand the mechanism of muscle growth,determination of meat quality and practice of molecular breeding.

关 键 词： 猪 肌肉组织 差异表达基因 消减 文库 抑制性消减杂交

领　　域： [农业科学] [农业科学]