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目的:应用酵母双杂交方法,寻找可能与Rab39A相互作用的蛋白。方法:采用醋酸锂转化法将诱饵蛋白Rab39A转入AH109酵母菌,Western blotting检测蛋白表达情况,检测诱饵载体有无毒性和自激活能力,筛选出的蛋白通过免疫荧光法进一步验证。结果:获得12个与Rab39A相互作用的候选蛋白,免疫荧光法证明Rab39A与ATRN、TIFA、ITGB1蛋白确实存在互作关系。结论:筛选出的蛋白主要与细胞生长、肿瘤细胞形成有关,为Rab39A功能的研究提供了重要线索。
OBJECTIVE: To find a possible protein interacting with Rab39A by yeast two-hybrid method. Methods: The bait protein Rab39A was transformed into AH109 yeast by lithium acetate conversion method. The protein expression was detected by Western blotting. The bait vector was tested for its toxicity and self-activation ability. The selected proteins were further verified by immunofluorescence. RESULTS: Twelve candidate proteins that interacted with Rab39A were obtained. Immunofluorescence showed that Rab39A interacted with ATRN, TIFA and ITGB1 proteins. Conclusion: The selected proteins are mainly related to cell growth and tumor cell formation, which provide important clues for the study of Rab39A function.