摘要
以四川省金堂黑山羊卵巢为材料,从其卵巢组织中提取总RNA,应用SMART^TM cDNA library construction技术,构建以真核表达载体pGADT7-Rec为基础的黑山羊卵巢酵母双杂交cDNA文库。结果表明,提取的RNA的A_260/A_280为1.95,28S与18S带浓度及亮度比值约为2。文库dscDNA弥散较长,分布在0.2～3kb,成瀑布条带状,而且在接近1kb的区域有密集的条带,为高丰度表达基因。根据生长菌落计数,共获得1.6×10~6个转化子,文库插入片段大小约为0.1～2kb。成功构建了金堂黑山羊卵巢酵母双杂交cDNA文库,为黑山羊多胎相关因子的基因筛选奠定了基础。
Taking the ovary of Jintang black goats as material, the yeast two-hybrid cDNA library of goat ovary was constructed by the technologies of CLONTECH SMART and homologous recombination. The results showed that 1.6 × 10^6 recombinants were obtained from the goat ovary cDNA library and the insert fragments of recombinants were demonstrated to distribute among 0.1-2kb in size. The yeast two-hybrid cDNA library of goat ovary in yeast cell AH109 was successfully constructed.
出处
《草业与畜牧》
2008年第3期41-43,53,共4页
Pruataculture & Animal Husbandry
基金
四川省应用基础项目(黑山羊多胎性状的分子机理研究)
四川省科技攻关计划项目
