多发性硬化症相关蛋白Mayven全长及结构域在酵母双杂交系统中的表达及对报告基因激活作用的检测

Assay of the expression and self-activition function of full length and different fragments of mayven gene in yeast two-hybrid system

目的探索Mayven在多发性硬化症病程中的作用以及寻找能与之发生相互作用的蛋白。方法通过PCR扩增得到Mayven基因全长P1及4个不同长度的Mayven基因片段，构建出包含Mayven全长P1及4个不同结构域片段的酵母双杂交诱饵载体，然后将其导入酵母菌株MAV203中，并检测其表达产物在酵母细胞中对报告基因的激活作用。结果实验结果表明全长P1、片段P3、P4不能激活报告基因HIS3和LacZ的表达，而片段P7和P8可以激活报告基因的表达，Mayven的C末端可能存在一个转录激活区。结论提示在筛选相互作用蛋白时可以使用全长P1及片段P3、P4，但转录激活是否是Mayven的生理功能仍有待进一步研究确证。

Objective To identify the mayven-interacting proteins and study the effect of mayven on multiple sclerosis pathogenesis. Methods The full length and four different fragments of mayven gene were amplified by polymerase chain reaction （PCR） with specific primers and were cloned into the yeast expression vector pDBLeu. Then, the recombinant plasmids were transformed into MAV203 yeast strain. The autonomous activation of their expression products was detected. Results The yeast expression vectors of mayven, which include a full-length and four different fragments, were constructed successfully. Full length P1, fragments P3 and P4 have no effect on the expression of HIS3 and LacZ gene, but fragments P7 and P8 do. The C-terminal of Mayven gene may contain a transcription activation domain. Conclusion Full length P1, fragments P3 and P4 of the mayven gene can be used to screen the mayven-interacting proteins, but whether Mayven has transcriptional activation activity need to be studied.