Stathmin基因磷酸化位点突变真核表达载体构建

发布时间：2018-04-25 11:08

  本文选题：Stathmin + 磷酸化　； 参考：《新乡医学院学报》2016年11期

【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。
[Abstract]:Objective to construct Stathmin gene phosphorylation site mutant vector and provide experimental basis for further study of Stathmin phosphorylation function. Methods four serine phosphorylation sites Ser16 Ser25Ser25Ser38 Ser63 in human Stathmin gene fragments were mutated to alanine which could not be phosphorylated by polymerase chain reaction (PCR). The eukaryotic expression vector pcDNA3.1 was inserted into the eukaryotic expression vector, and the recombinant vector PC DNA3.1/Stathmin Ser16% APC DNA3.1/Stathmin Ser25% APC DNA3.1/Stathmin Ser38% APC DNA3.1/Stathmin Ser63 / A was constructed, and the gene was sequenced. Results the recombinant vector was identified by gene sequencing. The results showed that the Ser16Con Ser25Ser25Ser38 Ser63 gene was mutated to alanine in the Stathmin gene. Conclusion the mutant vector of Stathmin phosphorylation site was successfully constructed, which laid a foundation for further study on the mechanism and function of Stathmin phosphorylation.
【作者单位】： 第四军医大学唐都医院临床实验与检验科;
【基金】：国家自然科学基金资助项目(编号:81372836)
【分类号】：R73-36
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本文编号：1801060