香蕉凝集素BanLec基因转化毕赤酵母及转化子的鉴定

BanLec Transformed into Pichia pastorisand and Identification of the Transformants

以pMD-BanLec质粒为模板扩增BanLec基因片段,该片段经EcoR I/XbaI双酶切后定向克隆到同样经EcoRI/XbaI双酶切的pPICZα表达载体上。将连接产物转化感受态DH5α,用低盐Zeocin抗性LB固体培养基筛选阳性克隆菌落。将重组质粒电转化毕赤酵母菌GS115后,通过PCR鉴定目的基因整合入酵母菌的基因组中,将有助于进一步研究BanLec蛋白的表达,为探讨香蕉凝集素的活性及生化功能等奠定基础。

BanLec fragment was amplified by PCR from pMD-BanLec vector,then the product was digested by EcoR I/Xba I to obtain BanLec fragment;the fragment was cloned in pPICZαto form pPICZα-BanLec recombinant expression vector.After E.coli DH5αwas transformated by pPICZα-BanLec vector,positive cloning strains were selected by low salt LB medium with Zeocin and identificated by restriction analysis by EcoR I/Xba I.The PCR result showed that recombinant plasmid pPICZα-BanLec was transformed into GSl15 by electroporation.The results laid a foundation for further study on the protein expression of BanLec and were helpful for investigating the activities or other physiological functions of BanLec.