摘要
目的 :寻找与 Cp G免疫激活序列 (imm unostimulatory sequence,ISS)有选择性相互作用的蛋白 ,以进一步探讨其分子作用机制。 方法 :采用酵母单杂交系统 ,以 4重复的 ISS为“诱饵”,对人骨髓细胞 c DNA文库进行初步筛选 ,并对部分阳性克隆以电泳迁移率变动分析 (EMSA )进行验证。结果 :已获得 4个双阳性克隆 ,其中两个编码功能未知的蛋白 ,而另外两个均属抗体轻链 ,分别与抗 DNA抗体和抗乙肝表面抗原抗体 (HBs Ab)高度同源 ;EMSA发现 HBs Ab可在偏酸 p H条件下 (p H6 .4和 5 .8时 )特异性地与含 ISS的寡核苷酸 (Cp G- ODN)结合。 结论 :HBs Ab可与
Objective: To screen out DNA binding proteins specifically recognizing CpG immunostimulatory sequence (ISS) for further investigating the molecular mechanisms of ISS. Methods: Yeast one hybrid system was adapted in screening a human bone marrow cDNA library using 4 copies of ISS as bait. The ISS binding activity of the positive clone was confirmed by electrophoretic mobility shift assay (EMSA). Results: Four dual positive colonies were obtained, two of them encoded proteins with unknown functions. The other 2 encoded light chains of immunoglobulin with amino sequences homology to anti DNA Ab and HBsAb respectively. EMSA showed HBsAb specifically bound to CpG ODN at pH6.4 and pH 5.8. Conclusion: HBsAb may have ISS specific DNA binding activity.
出处
《第二军医大学学报》
CAS
CSCD
北大核心
2001年第6期542-545,T002,共5页
Academic Journal of Second Military Medical University
基金
国家"8 63"计划资助项目 (10 1-0 6-0 5 0 4) .
