Screening out CpG immunostimulatory sequence-specific DNA-binding proteins with Yeast-one-hybrid system
- VernacularTitle:应用酵母单杂交系统筛选CpG免疫激活序列的特异性结合蛋白
- Author:
Zhenlin HU
;
Shuhan SUN
;
Jianxin DAI
;
Fengjuan ZHOU
;
- Publication Type:Journal Article
- Keywords:
CpG immunostimulatory sequence;
Yeast one hybrid system;
immunoglobulin
- From:
Academic Journal of Second Military Medical University
1985;0(06):-
- CountryChina
- Language:Chinese
-
Abstract:
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.
