SSRP1基因的克隆及其参与DNA损伤应答的初步研究  

Cloning of SSRP1 and its function analysis in response to DNA damage

作　　者：江洁[1] 洪泽辉[2] 

机构地区：[1]南京市市级机关医院内科 [2]东南大学医学院遗传与发育生物学系

出　　处：《东南大学学报（医学版）》2011年第6期865-868,共4页Journal of Southeast University(Medical Science Edition)

基　　金：国家自然科学基金资助项目(90919007);教育部新世纪优秀人才支持计划资助项目(NCET-09-0287)

摘　　要：目的:研究组蛋白伴侣分子SSRP1对DNA损伤的应答情况。方法:从HeLa细胞扩增SSRP1 cDNA,并克隆到EGFP载体上。EGFP-SSRP1在细胞内表达,用405 nm激光刺激诱导DNA损伤,随后观察EGFP-SSRP1的聚集情况。结果:经酶切和测序鉴定,我们成功构建了EGFP-SSRP1表达载体。EGFP-SSRP1在405 nm激光刺激后迅速聚集到激光照射位点。结论:我们的结果直接证实了SSRP1快速参与DNA损伤应答。Objective： To investigate the function of histone shaperone SSRP1 in response to DNA damage.Methods： SSRP1 gene was amplified from HeLa cells,cells plated in glass-bottomed and cloned to EGFP vector.EGFP-SSRP1 was transiently transfected into HeLa dishes.Cells expressed EGFP-SSRP1 was micro-irradiated with a 405 nm pulse laser along a user-defined path.The response of SSRP1 to DNA damage was measured by a confocal scanning laser microscopy system.Results： The EGFP-SSRP1 vector was identified and confirmed by enzyme digestion and sequencing.SSRP1 protein was recruited to DNA damage sites induced by 405 nm laser.Conclusion： SSRP1 is associated with DNA damage response.

关 键 词：基因克隆 DNA损伤 聚集 

分 类 号：Q785[生物学—分子生物学]