摘要
以花生品系E12幼苗为材料分离mRNA,利用SMART（Switching Mechanism at 5＇end ofRNA Transcript）技术合成双链cDNA。经限制性内切酶SfiⅠ酶切,将经过分级分离得到的cD-NA连接到改造的质粒载体pBluescriptⅡSK,构建了花生幼苗cDNA文库。经检测,所构建的文库的滴度为1.1×106cfu/mL,重组率为93.4%,插入片段大小为750-2000bp。
In order to study the novel genes of peanut, a cDNA library from seedlings was constructed. Total RNA was extracted from E12 peanut seedlings and mRNA was purified. Double strand cDNA was synthesized by SMART method, the ds cDNA fragments were ligated into the pBluescriptⅡSK vector. The recombinant plasmids were transformed into the E. coli, a cDNA library of peanut seedlings was successfully constructed. The titer of the cDNA library was estimated as 1.1×10^6cfu/mL; the percentage of recombination was 93.4%. PCR results showed that the inserts varied from 0.75 to 2.0 kb.
出处
《花生学报》
2010年第2期11-15,共5页
Journal of Peanut Science
基金
现代农业产业技术体系建设专项资金(nycytx-19)
国家高技术研究发展计划项目(2007AA10Z189
2006AA10A114)
国家科技支撑计划(2008BAD97B04)
