山核桃脂肪代谢相关cDNA文库的构建  

Construction of a fat metabolism-related cDNA library in Carya cathayensis

作　　者：黄银芝[1] 周秦[1] 黄有军[1] 曾燕如[1] 

机构地区：[1]浙江农林大学亚热带森林培育国家重点实验室培育基地,浙江临安311300

出　　处：《浙江农林大学学报》2011年第1期80-85,共6页Journal of Zhejiang A&F University

基　　金：浙江省自然科学基金资助项目(Y306373);浙江省森林培育重中之重学科开放基金资助项目(200609)

摘　　要：山核桃Carya cathayensis种仁含油量较高,油质多为不饱和脂肪酸。在差异显示反转录-聚合酶链式反应（DDRT-PCR）方法研究山核桃果实成熟时期基因差异表达的基础上,采用SMART（switching mechanism at 5’end of RNAtranscript）技术构建了山核桃脂肪代谢时期的互补脱氧核糖核酸（cDNA）文库。未扩增文库的滴度为5.0×105个噬菌斑形成单位（pfu）.mL-1,插入片段大小为0.7~2.5 kb,70.0%的片段超过1.0 kb;扩增文库的滴度为5.0×109pfu.mL-1,cDNA片段大小均在500 bp以上,重组率达94.5%,66.0%的片段超过1.0 kb。文库的构建为研究山核桃脂肪代谢机制、调控网络及相关基因的发现提供了条件。Carya cathayensis is a species with a high oil content in its nuts.In addition,its oil is mostly unsaturated fatty acids.Based on differential display of genes during fruit maturity studied by DDRT-PCR,a fat metabolism-related cDNA library was constructed by SMART.Titering the unamplified library showed a titer of 5.0 × 105 pfu·mL-1,an insert size between 0.7 and 2.5 kb and 70.0% of inserts larger than 1.0 kb;and the amplified library had a titer of 5.0 × 109 pfu·mL-1,all of the inserts larger than 500 bp,and a recombination rate of 94.5% with 66.0% of the inserts longer than 1 000 bp.This work has laid a foundation for studies of fat-metabolizing mechanism and regulatory network as well as discovery of relevant genes.[Ch,7 fig.19 ref.]

关 键 词：经济林学 山核桃 CDNA文库 SMART 脂肪代谢 

分 类 号：S718.46[农业科学—林学]