应用酵母双杂交筛选与hSOX4相互作用的蛋白质

Screening of proteins interacting with hSOX4 by yeast two-hybrid

目的:寻找与hSOX4相互作用的蛋白质,为该基因的功能研究提供新的线索.方法:采用酵母双杂交方法,以hSOX4的第1～133位氨基酸SOX4(1-133)和第130～380位氨基酸SOX4(130-380)分别作为诱饵筛选人乳腺文库,经过重复验证排除假阳性以确定阳性克隆.结果:以SOX4(1-133)作为诱饵最终筛出了4个阳性克隆,经测序及生物信息学分析,这4个克隆为同一基因来源:UBE2I(ubiquitin-conjugating enzyme E2I, Ubc9).以SOX4(130-380)为诱饵未能筛出阳性克隆.结论:Ubc9能与SOX4(1-133)发生相互作用,它们的相互作用可能与SOX4的转录调控有关.

Objective： To look for proteins interacting with human SOX4. Methods： Yeast two-hybrid was applied to screen human breast cDNA library. Two fragments of hSOX4, SOX4（1-133aa） and SOX4（130-380aa）, were used as the baits respectively. Results： hUbc9, a class E2 conjugating enzyme of small ubiquitin-related modifier （SUMO）-1, was identified to interact with SOX4 （1-133aa） and no positive clones were identified when the bait was SOX4 （130-380aa）. Conclusion： Ubc9 can interact with SOX4 （1-133）. Their interaction may suggest a new mechanism to regulate the transcriptional activity of SOX4.