[1]杨　铭,林　芳,和　婷,等.Stathmin基因磷酸化位点突变真核表达载体构建[J].新乡医学院学报,2016,33(11):941-944.[doi:10.7683/xxyxyxb.2016.11.002]
　YANG Ming,LIN Fang,HE Ting,et al.Construction of Stathmin gene phosphorylation site mutation eukaryotic expression vector[J].Journal of Xinxiang Medical University,2016,33(11):941-944.[doi:10.7683/xxyxyxb.2016.11.002]
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Stathmin基因磷酸化位点突变真核表达载体构建
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《新乡医学院学报》[ISSN:1004-7239/CN:41-1186/R]

卷:
33
期数:
2016年11
页码:
941-944
栏目:
基础研究
出版日期:
2016-11-05

文章信息/Info

Title:
Construction of Stathmin gene phosphorylation site mutation eukaryotic expression vector
作者:
杨　铭; 林　芳; 和　婷; 王　琳; 汪定成; 董　轲; 张惠中
（第四军医大学唐都医院临床实验与检验科，陕西　西安　710038）
Author(s):
YANG Ming; LIN Fang; HE Ting; WANG Lin; WANG Ding-cheng; DONG Ke; ZHANG Hui-zhong
(Central Laboratory of Tangdu Hospital,the Fourth Military Medical University,Xi′an 710038，Shaanxi Province,China)
关键词:
Stathmin; 磷酸化; 点突变; 载体构建
Keywords:
Stathmin; phosphorylation; point mutation; vector construction
分类号:
R363
DOI:
10.7683/xxyxyxb.2016.11.002
文献标志码:
A
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pcDNA3.1，分别构建重组载体pcDNA3.1/Stathmin Ser16/A、pcDNA3.1/Stathmin Ser25/A、pcDNA3.1/Stathmin Ser38/A、pcDNA3.1/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。
Abstract:
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 Ser16，Ser25，Ser38 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 Ser16/A,pcDNA3.1/Stathmin Ser25/A,pcDNA3.1/Stathmin Ser38/A and pcDNA3.1/Stathmin Ser63/A were constructed,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.

参考文献/References:

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更新日期/Last Update: 2016-11-05