【摘要】 了解肌肉组织和肝脏组织的基因类型,构建鸡肌肉组织和肝脏组织的功能基因表达谱在动物遗传育种中具有重要的意义。为比较基因组学的研究、功能基因组学的研究、QTLs定位等的动物分子育种做前期基础科研工作,研究以丝毛乌骨鸡(Gallusgallus)的胸肌和肝脏为实验材料,以LambdaZAPⅡ为载体,构建了胸肌和肝脏的cDNA文库。同时,对这两个cDNA文库进行了噬菌体PCR鉴定和所提质粒的Eco RⅠ和XhoⅠ的双酶切鉴定。结果表明,胸肌和肝脏mRNA的OD260/OD280 分别为1.92和1.90。胸肌cDNA文库的滴度在3.6×107 pfu/mL以上,重组率92%以上,插入片段平均大于1.5kb。肝脏cDNA文库的滴度在3.4×107 pfu/mL以上,重组率90%以上,插入片段平均大于1.4kb。更多还原

【Abstract】 More and more animal breeding scientists are interested in chicken economic trait research. Most of these traits are related to different gene expression. So understanding the expressed genes and constructing the known gene expressed profile from muscle tissue and liver tissue of chicken are very important in animal breeding and genetics research. In order to prepare for the future comparative genomics research, functional genomics research, QTLs mapping, and animal molecular breeding etc, we have successfully constructed a breast muscle tissue cDNA libraries and a liver tissue cDNA libraries of Silkie Fowl (Gallus gallus ) in the study. The vector of cDNA used was Lambda ZAP II. At the same time, both cDNA libraries’ quality were identified by the PCR for phage, the enzyme of EcoRⅠand XhoⅠ digested for plasmid and other methods. The results showed that the OD260/OD280 of mRNA from breast muscle and liver were 1.92 and 1.90 respectively, the titer of the breast library was 3.6×107 pfu/mL,the recombinant percentage was 92%,and the average length of inserted cDNA fragments was longer than1.5 kb. The titer of the liver cDNA library was 3.4×107 pfu/mL,the recombinant percentage was 90%,and the average length of inserted fragment cDNA was longer than 1.4 kb.更多还原