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目的 构建干扰素诱导蛋白P5 6及其各种缺失突变体的酵母表达质粒。方法 干扰素诱导HT10 80 ,提取mRNA、RT -PCR获取P5 6全长及各种缺失突变体的cDNA ,以酵母表达质粒pGADT7为载体 ,构建重组质粒。结果 诱导的HT10 80中抽提出P5 6的mRNA、RT -PCR扩增出P5 6全长cDNA片段 ,大小与预期值一致约 15 0 0bp ,以P5 6全长cDNA为模板PCR获得P5 6各种缺失突变体并插入pGADT7;正确构建了各种pGADT7 P5 6酵母表达重组质粒。这些质粒DNA测序结果与理论值一致。结论 P5 6全长及各种缺失突变体酵母表达质粒的成功构建 ,为进一步研究P5 6功能奠定了基础
Objective To construct a yeast expression plasmid of interferon - inducible protein P5 6 and its various deletion mutants. Methods Interferon induced HT1080, mRNA was extracted, cDNA of full length P5 6 and various deletion mutants were obtained by RT-PCR. The recombinant plasmid was constructed by using yeast expression plasmid pGADT7 as a vector. Results The mRNA of P5 6 was extracted from HT10 80 induced by RT-PCR, and the full-length cDNA of P5 6 was amplified by RT-PCR. The full-length cDNA of P5 6 was approximately identical to the expected value of 150 bp. The mutant was deleted and inserted into pGADT7. Various pGADT7 P5 6 yeast expression recombinant plasmids were constructed correctly. These plasmid DNA sequencing results consistent with the theoretical value. Conclusion The successful construction of P5 6 full-length and various deletion mutant yeast expression plasmids laid the foundation for further study of P5 6 function