大鼠脑源性神经营养因子基因T载体的构建及鉴定

Construction and identification of brain derived neurotrophic factor gene cloned into a T-vector in rat

中文关键词:  脑源性神经营养因子  T载体  基因克隆

英文关键词:brain derived neurotrophic factor  T-vector  Gene cloning

中文摘要:

  目的：探讨利用T载体完成大鼠脑源性神经营养因子（BDNF）聚合酶链式反应（polymemse chain reaction，PCR）产物克隆的可行性。方法：提取大鼠脑组织总RNA，利用M-MLV逆转录酶及oligo-dT引物逆转录合成cDNA第一条链，再利用exTaq酶及BDNF上、下游引物扩增出BDNF cDNA．最后利用Buffer Ⅰ连接液将BDNF基因克隆入T载体用于测序及下一步克隆。结果：成功构建了BDNF基因T载体．经双脱氧链终止法测序证实为大鼠BDNF基因。结论：利用T载体克隆BDNF基因PCR产物简便、快捷，成功率高。

英文摘要:

  Objective:To construct a T-vector cloned with a BDNF gene of a SD rat.Method:The total RNA of the brain of a SD rat were extracted,the first strand cDNA was synthesized using M-MLV reverse transcriptase and oligo-dT primers,and then BDNF cDNA was amplifie