1.Screening of protein interacting with the transcript of UL128 gene showed two protein patterns by yeast two-hybrid from human fetus brain cDNA library
Gaowei REN ; Xin CUI ; Yanping MA ; Ying QI ; Qiang RUAN ; Zhengrong SUN
Chinese Journal of Microbiology and Immunology 2010;30(8):737-742
Objective Using yeast two-hybrid system to screen the proteins which can interact with the human cytomegalovirus (HCMV) UL128 which have two difference transcription structure from human fetus brain cDNA library, and compare the difference with structure and function of interacting proteins. Methods Two fragments of UL128 were amplified by 3'RACE and 5'RACE technology, the length are 519 bp and 642 bp, respectively. The "bait plasmid" (named as pGBKT7-UL128-519 bp and pGBKT7-UL128-642 bp) was constructed successfully. Using pGBKT7-UL128-519 bp and pGBKT7-UL128-642 bp as a bait, a human fetus brain cDNA was screened and the proteins interacting with UL128-519 bp and UL128-642 bp encoded protein were searched, and the positive clones were sequenced and analyzed by bioinformatic methods. Results EFEMP2 interacting with HCMV UL128-519 bp were identified, THY-1 interacting with HCMV UL128-642 bp were identified. Conclusion EFEMP2 and THY-1 proteins interacting with HCMV UL128-519 bp and UL128-642 bp in human fetus brain cDNA library were successfully screened, but same proteins weren't found from the proteins interacting with UL128-519 bp and UL128-642 bp protein, UL128-519 bp and UL128-642 bp protein may be play an different effect in the process of infect by HCMV.
2.Screening of Protein Interacting with HCMV UL130 Protein by Yeast Two Hybrid from Human Fetus Brain cDNA Library
Gaowei REN ; Xin CUI ; Ying QI ; Yanpin MA ; Qiang RUAN ; Zhengrong SUN
Journal of China Medical University 2010;(6):413-416
Objective To screen the human proteins interacting with human cytomegalovirus(HCMV)UL130 from human fetus brain cDNA library by GAL4 two-hybrid system 3 technique and analyze the corresponding coding sequences.Methods The "bait plasmid"(named as pGBKT7-UL130)was constructed.By using HCMV UL130 as the bait,a human fetus brain cDNA library was screened and the proteins interacting with UL130 protein were searched.The positive clones were sequenced and analyzed by bioinformatic methods.Results Nine clones interacting with HCMV UL130 were identified,two of them were synaptosome-associated protein(SNAP).Conclusion Some proteins interacting with HCMV UL130 in human fetus brain cDNA library were successfully screened.SNAP might play an important role in HCMV infection pathogenesis.
3.It is necessary to quickly establish the reference method and the traceability cyctem for GA in China
Rui ZHANG ; Wenhua REN ; Huaian MA ; Mo WANG ; Gaowei FAN ; Qingtao WANG
Chinese Journal of Laboratory Medicine 2017;40(10):741-743
Diagnosis and treatment of diabetes mellitus depend on accurate monitor of blood glucose of laboratory.Glycated albumin,the short-term indicator of blood glucose control, has special advantage and role in the monitor of diabetes mellitus and has been payed more and more attention.The standardization work of determination of glycated albumin at home and abroad is still in the primary stage at present.The consistency and accuracy of measurement results for glycated albumin is needed to be solved urgently.