利用FRET技术研究小鼠PU.1和KLF7蛋白相互作用  

The Study of Mouse PU.1 and KLF7 Protein Interaction by FRET Technology

作　　者：李雯 邵淑丽[1] 张珍珠[1] 谭茗 金钊 李金炜 孙婴宁[1] LI Wen;SHAO Shuli;ZHANG Zhenzhu;TAN Ming;JIN Zhao;LI Jinwei;SUN Yingning(College of Life Science and Agriculture and Forestry,Qiqihar University,Qiqihar 161006,China)

机构地区：[1]齐齐哈尔大学生命科学与农林学院,齐齐哈尔161006

出　　处：《中国细胞生物学学报》2021年第9期1729-1736,共8页Chinese Journal of Cell Biology

基　　金：国家自然科学基金(批准号:31402061);黑龙江省自然科学基金(批准号:YQ2019C025);黑龙江省省属高等学校基本科研业务费科研项目(植物性食品加工技术特色学科专项)(批准号:YSTSXK201875)资助的课题。

摘　　要：PU.1和KLF7基因在多个组织中的表达具有相关性。在前脂肪细胞分化过程中,两个转录因子都可以通过抑制PPARγ的表达而抑制脂肪形成。然而两者是否存在蛋白水平相互作用还不清楚。为了验证PU.1和KLF7在前脂肪细胞中的关系,该研究构建了重组表达载体pECFP-N1-PU.1和pEYFP-N1-KLF7,并瞬时转染到3T3-L1前脂肪细胞系中。利用激光共聚焦显微镜,观察到PU.1和KLF7蛋白共定位于3T3-L1细胞核中。荧光共振能量转移(fluorescence resonance energy transfer,FRET)实验分析显示,PU.1和KLF7之间有明显的能量共振转移。实验结果证实,PU.1和KLF7之间存在直接的相互作用关系,为脂肪生成的调控机制研究提供新思路。The expressions of PU.1 and KLF7 genes are significantly positively correlated in multiple tissues.Both the two transcription factors inhibit adipogenesis through repressing PPARγ expression during preadipocyte differentiation.However,little is known about their interaction.To verify the relationship between PU.1 and KLF7 in preadipocytes,in the present study,recombinant overexpression vector pECFP-PU.1 and pEYFP-KLF7 were constructed and transfected into 3T3-L1 cell lines.Laser confocal microscope was used to observe their co-localization in the nuclei of 3T3-L1 cell lines.The FRET (fluorescence resonance energy transfer) analysis revealed a significant energy resonance transfer between PU.1 and KLF7.These results confirm that there is a direct interaction between PU.1 and KLF7,which provides a new idea for the mechanism of regulating adipogenesis.

关 键 词：荧光共振能量转移 PU.1 KLF7 3T3-L1 

分 类 号：Q756[生物学—分子生物学]