本文選題：Stathmin + 磷酸化　； 參考：《新鄉(xiāng)醫(yī)學(xué)院學(xué)報(bào)》2016年11期

【摘要】：目的構(gòu)建Stathmin基因磷酸化位點(diǎn)突變載體,為Stathmin磷酸化功能的進(jìn)一步研究提供實(shí)驗(yàn)基礎(chǔ)。方法通過設(shè)計(jì)磷酸化位點(diǎn)突變引物,運(yùn)用聚合酶鏈反應(yīng)分別將人Stathmin基因片段中4個(gè)絲氨酸磷酸化位點(diǎn)Ser16、Ser25、Ser38、Ser63突變?yōu)椴荒苓M(jìn)行磷酸化的丙氨酸,插入真核表達(dá)載體pc DNA3.1,分別構(gòu)建重組載體pc DNA3.1/Stathmin Ser16/A、pc DNA3.1/Stathmin Ser25/A、pc DNA3.1/Stathmin Ser38/A、pc DNA3.1/Stathmin Ser63/A,并進(jìn)行基因測序鑒定。結(jié)果基因測序鑒定重組載體結(jié)果表明,分別將Stathmin基因中Ser16、Ser25、Ser38、Ser63突變?yōu)楸彼�。結(jié)論成功構(gòu)建了Stathmin磷酸化位點(diǎn)突變載體,為進(jìn)一步研究Stathmin磷酸化的作用機(jī)制和功能奠定了基礎(chǔ)。
[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.
【作者單位】： 第四軍醫(yī)大學(xué)唐都醫(yī)院臨床實(shí)驗(yàn)與檢驗(yàn)科;
【基金】：國家自然科學(xué)基金資助項(xiàng)目(編號:81372836)
【分類號】：R73-36
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本文編號：1801060