猪杂种一代与亲本肌肉组织间正反向消减cDNA文库的构建

Establishment of Forward-and Reverse-Subtracted cDNA Librariesbetween Muscle Tissue of F1 and Parental Generation Pigs

应用抑制消减杂交技术成功构建了杂种一代梅大猪、亲代大白猪的背最长肌正反向消减cDNA文库。以βactin为指标检测2个文库的消减效率分别为215和210倍。从相应的质粒文库中分别获取了700个和623个有效阳性克隆,PCR检测插入片段主要分布在150~600 bp之间。亲本与子代肌肉组织间消减cDNA文库的构建为进一步分离、鉴定与杂种优势相关的基因奠定了基础。

Forward-and reverse-subtractive cDNA libraries between Longissimus Dorsi of Largewhite and F1 of Meishan×Largewhite pigs have been successfully generated by means of suppression subtractive hybridization(SSH), and both of them were subject to the determination of subtraction efficiency that proved to be approximately 2^(15)-and 2^(10)-fold indicated by β-actin as an index. PCR analysis of the total of 700 and 623 positive clones obtained from the corresponding plasmid libraries demonstrated that the cDNA inserts range in size largely from 150 to 600bp. It is safe to say that the construction of forward-and reverse-subtractive cDNA libraries between muscle tissue of parents and progenies lays groundwork for isolating and identifying genes with respect to heterosis.