%0 Journal Article
%T 小鼠PICK1与Orai1蛋白质间的相互作用研究<br>Research on Interaction between PICK1 and Orai1 of Mouse
%A 蔡佩玲
%A 程丽佳
%A 王婷
%J 四川大学学报 (自然科学版)
%D 2015
%X 从小鼠脑组织中提取总RNA，RT PCR扩增PICK1及Orai1的CDS序列，构建原核表达载体GST PICK1及真核表达载体myc Orai1，重组质粒经？？Sal？？Ⅰ和？？Not？？Ⅰ双酶切鉴定后测序. 正确的质粒myc Orai1在HEK293细胞中表达收获过表达蛋白，GST PICK1进行原核表达并获得融合蛋白. 将纯化后的GST PICK1蛋白条带切下，进行蛋白质液相质谱分析. 后将myc Orai1蛋白与GST PICK1蛋白混合，进行GST pulldown实验，western blot检测到myc Orai1的条带. 结果表明PICK1与Orai1在体外有相互作用.<br>Total RNA of mice brain was extracted, CDS fragment of PICK1 and Orai1 was obtained by RT PCR, CDS were respectively sub cloned into pGEX 4T 2 with GST tag to generate GST PICK1 and sub cloned into pRK5 with myc tag to get myc Orai1, and recombinants were sequenced after confirmation by digested with ？？Sal？？Ⅰ and ？？Not？？Ⅰ. Correct myc Orai1 vector was expressed in HEK293 cells, GST PICK1 vector was expressed in BL21 competent cells. GST PICK1 fusion proteins were analyzed with protein liquid mass spectrometry after being purified and cut SDS PAGE gel of the target band. And then GST PICK1 fusion protein and myc Orai1 fusion protein were mixed to apply GST pulldown experiment, and myc Orai1 band was detected by western blot. Result indicates PICK1 and Orai1 interacted ？？in Vitro.？
%K PICK1 Orai1 GST Pulldown 相互作用&lt
%K br&gt
%K PICK1 Orai1 GST pulldown Interaction
%U http://science.ijournals.cn/jsunature_cn/ch/reader/view_abstract.aspx?file_no=20153039&flag=1