作　　者： ; ; ;

机构地区： 福建农林大学

出　　处： 《花生学报》 2007年第2期1-5,共5页

摘　　要： 以花生品种闽花5号各发育时期的种子为材料分离mRNA，利用SMART技术合成双链eDNA。利用限制性内切酶sfiⅠ酶切。将经过分级分离得到的cDNA连接到质粒载体pDNR-LIB，成功构建花生种子全长cDNA文库。将所得到初级文库扩增后进行保存，检测扩增文库滴度为1．7×10^9efu／mL。PCR鉴定重组子，发现重组率接近100N，插入片段集中分布在0．5～2．0kb。插入片段的平均大小在1000bp左右，这说明该文库既可以满足低丰度基因的筛选，也可保证获得全长cDNA。 In order to study the novel genes of peanut seeds, a full-length cDNA library from peanut seeds was constructed, mRNA from peanut seeds of different developmental stages was purified, Double strand cDNA was synthesized by SMART method, the ds cDNA fragments were ligated to the pDNR-LIB vector. The recombinant plasmids were transformed into the E. coli, a cDNA library of peanut seeds was successfully constructed. The fulllength cDNA library was stocked after amplification, the titer of the cDNA library was estimated as 1.7× 10^9 cfu/ mL, PCR results showed that the inserts varied from 0.5 to 2.0kb with average size was larger than 1000bp. It indicated that this library could be used for full-length genes screening and cloning of low abundance genes.

关 键 词： 花生 种子 文库 技术

领　　域： [农业科学] [生物学]