摘要
目的:探讨黄瓜性别相关基因ERAF17在黄瓜性别调控过程中的作用。方法:通过RT-PCR法克隆黄瓜性别相关基因ERAF17,利用分子克隆技术将目的片段连入植物化学诱导表达载体pER8,选择部分ERAF17序列构建含有目的片段发卡结构的沉默载体。构建原核表达载体pET-28a-ERAF17在大肠杆菌中诱导表达ERAF17蛋白并进行纯化。结果:成功构建了植物化学诱导表达载体pER8-ERAF17和含有目的片段发卡结构的沉默载体pMBW330-ERAF17-hp,在含有pET-28a-ERAF17原核表达载体的大肠杆菌中加入IPTG诱导3h后ERAF17表达量最高,通过NT-agerose介质成功纯化出ERAF17的编码蛋白。结论:所构建载体可用于ERAF17基因的功能研究。
Objective:Research on the gene function of cucumber sex determination related gene ERAF17.Method:The ERAF17 gene was constructed into chemically inducible vector pER8 and part sequence of ERAF17 was selected to form the hairpin structure by the molecular technology,prokaryotic expression vector pET-28a-ERAF17 was constructed and ERAF17 protein was expressed in Escherichia coli.Result:Plant expression vector pER8-ERAF17 and the hairpin structure pMBW330-ERAF17-hp were constructed,the highest accumulation of ERAF17 protein appeared after inducing by IPTG for 3 hours.Conclusion:These recombinant vectors can be used for researching gene function of ERAF17.
出处
《生物技术》
CAS
CSCD
北大核心
2012年第4期15-18,共4页
Biotechnology
