Stathmin基因磷酸化位点突变真核表达载体构建  

Construction of Stathmin gene phosphorylation site mutation eukaryotic expression vector

作　　者：杨铭[1] 林芳[1] 和婷[1] 王琳[1] 汪定成[1] 董轲[1] 张惠中[1] 

机构地区：[1]第四军医大学唐都医院临床实验与检验科,陕西西安710038

出　　处：《新乡医学院学报》2016年第11期941-944,共4页Journal of Xinxiang Medical University

基　　金：国家自然科学基金资助项目(编号:81372836)

摘　　要：目的构建Stathmin基因磷酸化位点突变载体,为Stathmin磷酸化功能的进一步研究提供实验基础。方法通过设计磷酸化位点突变引物,运用聚合酶链反应分别将人Stathmin基因片段中4个丝氨酸磷酸化位点Ser16、Ser25、Ser38、Ser63突变为不能进行磷酸化的丙氨酸,插入真核表达载体pc DNA3.1,分别构建重组载体pc DNA3.1/Stathmin Ser16/A、pc DNA3.1/Stathmin Ser25/A、pc DNA3.1/Stathmin Ser38/A、pc DNA3.1/Stathmin Ser63/A,并进行基因测序鉴定。结果基因测序鉴定重组载体结果表明,分别将Stathmin基因中Ser16、Ser25、Ser38、Ser63突变为丙氨酸。结论成功构建了Stathmin磷酸化位点突变载体,为进一步研究Stathmin磷酸化的作用机制和功能奠定了基础。Objective To construct the Stathmin gene phosphorylation site mutation eukaryotic expression vector, and to provide the experimental basis for the further research of Stathmin phosphorylation. Methods Stathmin phosphorylation site mutation primers were designed. The Serl6, Ser25 ,Set38 and Ser63 sites of human Stathmin gene were mutated to alanine by polymerase chain reaction,and cloned into the eukaryotic expression vector pcDNA3.1. The recombinant vectors pcDNA3.1/ Stathmin Serl6/A,pcDNA3.1/Stathmin Ser25/A,pcDNA3.1/Stathmin Ser38/A and peDNA3.1/Stathmin Ser63/A were con- structed, and the recombinant vectors were identified by gene sequencing. Results Gene sequencing results showed that the phosphorylation site of Stathmin gene was mutated to alanine successfully. Conclusion Stathmin phosphorylation site mutation vector is successfully constructed ,which provide the experimental basis for the further study on the mechanism and function of Stathmin phosphorylation.

关 键 词：STATHMIN 磷酸化 点突变 载体构建 

分 类 号：R363[医药卫生—病理学]