ABSTRACT
The yeast one hybrid system is the new type system deriving from the yeast two hybrid system to study the interaction between DNA and protein.The principle and the technical line of this system was briefly introduced,and the progress of this system on the research of plant transcription factor gene resistant to osmotic stress was particularly summarized,such as cloning the transcription factor genes of osmosis resistance type;confirming the interaction between known DNA and protein;making sure the localization of the confirmed DNA binding domain with the interaction;also validating the transcription activation of the transcription factor gene.Furthermore,the problems existing in this system on the research of plant transcription factor gene resistant to osmotic stress was analyzed,and the development prospect according to the research was forecasted.
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.