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目的:探讨TTF-1相关蛋白26(TAP26)磷酸化位点对SP-C基因表达调控的影响。方法:采用定点诱变PCR技术产生TAP26的4个磷酸化位点突变体TAP26(S48→A48)、TAP26(S66→A66)、TAP26(T167S168→V167A168)、TAP26(T219→V219);通过体外细胞共转染后荧光素酶报告基因活性值检测技术研究野生型TAP26及其4个突变体质粒对SP-C基因启动子表达活性的影响。结果:荧光素酶报告基因活性值检测结果显示,分别与共转染野生型TAP26和其它3个突变体质粒相比较,共转染TAP26(S66→A66)突变体质粒的荧光素酶活性值最低,经统计分析差异均有统计学意义(P<0.05,n=9)。结论:TAP26磷酸化对促进SP-C基因表达具有非常重要的作用,TAP-26的S66位点则是与TTF-1相互作用从而共同调控SP-C基因表达的一个关键PKC磷酸化位点。
Objective: To investigate the effect of TTAP-1-related protein 26 (TAP26) phosphorylation site on SP-C gene expression. Methods: TAP26 mutants TAP26 (S48 → A48), TAP26 (S66 → A66), TAP26 (T167S168 → V167A168) and TAP26 (T219 → V219) were generated by site-directed mutagenesis Effects of co-transfection of luciferase reporter gene activity assay on expression of SP-C promoter in wild-type TAP26 and its four mutant plasmids. Results: The results of luciferase reporter assay showed that luciferase activity of the co-transfected TAP26 (S66 → A66) mutant plasmid was the lowest, compared with the cotransfected wild-type TAP26 and the other three mutant plasmids respectively. The statistical analysis of differences were statistically significant (P <0.05, n = 9). CONCLUSION: TAP26 phosphorylation plays an important role in promoting the expression of SP-C gene. The S66 site of TAP-26 is a key PKC phosphorylation site that interacts with TTF-1 to regulate the expression of SP-C gene.