毛竹笋全长cDNA文库构建

Construction of a cDNA Library of Phyllostachs pubescens Using Gateway Technology

Gateway技术构建cDNA文库,利用了λ噬菌体的位点特异性重组,避免使用限制性内酶切割cDNA,能够解决常规方法构建cDNA文库的技术缺陷。应用Gateway技术构建毛竹笋cDNA文库,经检测cDNA入门文库的滴度达到1.7×106cfu/mL,文库总容量为8.5×106cfu,平均插入片段在1.0 kb以上。毛竹笋文库的构建为进一步克隆毛竹纤维化分子机理打下了基础。

A cDNA library was constructed based on the Gateway system that is a methed to construct a high quality cDNA library without the use of restriction enzyme for cloning. This is the first report that the Gateway system is used to construct a cDNA library for Phyllostachs pubescens. The entry cDNA library was constructed in this report has a high titer of 1.7 ×10^6cfu/mLand contain a total clones of 8. 5×10^6cfu/mL, with an average inserts size of about 1.0 kb. The construction of the cDNA library of bamboo shoots（Phyllostachs pubescens） which made the further step to fibrosis gene for cloning laid the groundwork for the molecular mechanism of fibrosis.