稻曲病菌cDNA文库的构建

 以稻曲病菌菌丝和薄壁分生孢子提取的总RNA为起始模板,利用SMART cDNA library construction kit构建稻曲病菌cDNA文库,并分析文库质量。结果显示,未扩增文库滴度9.03×10⁶pfu/mL,重组率为99.5%,扩增后文库滴度达5.52×10⁹pfu/mL。随机挑取10个噬菌斑转化成质粒,SfiⅠ酶切显示插入片段长度在400-2000bp之间;经测序得到10条表达序列标签(EST),GenBank数据库相似性联配结果显示6条EST具有同源序列。其中,克隆R2为乙酰胺酶同源物编码蛋白。

Construction of a cDNA library of the fungus Ustilaginoidea Wrens

A cDNA library driven from the total RNA extracted from mycelia and conidia of Ustilaginoidea virens was constructed using the SMART cDNA library construction kit.Then,the qualities of unamplified and amplified library were detected.The results showed that unamplified library consisted of 9.03×10⁶ pfu/mL independent clones,and the recombinant rate was about 99.5%.The titer of amplified cDNA library was 5.52×10⁹ pfu/mL.SfiⅠdigestion of ten random phage clones indicted that the sizes of cDNA included were among 400-2 000 bp.Sequencing of these ten clones revealed ten expressed sequence tags (ESTs),among which,six ESTs had homologous sequence in GenBank.EST R2 encodes a homologous protein of acetami-dase.