玉米纹枯病菌全长cDNA文库的构建与鉴定

Construction and characterization of a full-length c DNA library of Rhizoctonia solani

以玉米纹枯病菌菌丝为材料,利用In-Fusion SMARTer TM c DNA Lib Construction Kit构建玉米纹枯病菌全长c DNA文库。文库质量鉴定结果表明,文库滴度为1.2×106pfu·m L-1,重组率为99.2%,插入片段平均长度大于1.0 kb。随机挑取400个白色克隆进行测序,共获得329个高质量EST序列,经聚类拼接后得到250条unique EST,包括36条contigs和214条singletons。在Gen Bank进行Blastn与Blastx同源比对,有227条EST与已知核酸或蛋白有不同程度的同源性,占全部EST的90.8%,其余23条无任何同源性,占9.2%。

A full-length cDNA library was constructed from hypha of Rh＆octonia solani by using In-Fusion SMARTerTM cDNA Library Construction Kit. The constructed library had a high titer of 1.2×10^-pfu·mL^-1 and a 99.2% recombination rate, of which the average length of the inserted cDNA fragment was above 1.0 kb. The 329 high quality ESTs were obtained from 400 white clones picked randomly. After treatments of clustering and splicing, 250 unique ESTs were obtained, including 36 contings and 214 singlets. These unique ESTs were analyzed for sequence homology with Blastx and Blastn software against sequences in the GenBank. Out of the 250 unique ESTs , 227 ESTs that accounted for 90.8% of all the unigenes showed homology with the function known genes or proteins, whereas the rest that accounted for 9.2% has no homology.