Identification of the interactions between the truncated fragments of acrophage migration inhibitory factor and CD74 using yeast two-hybrid system
会议名称：《中国病理生理学会受体和信号转导专业委员会暨消化专业委员会联合学术会议》
会议日期：2008年
学科分类：0710[理学-生物学] 07[理学] 08[工学] 09[农学] 071007[理学-遗传学] 0901[农学-作物学] 0836[工学-生物工程] 090102[农学-作物遗传育种]
关 键 词：yeast two-hybrid system macrophage migration inhibitory factor CD74 protein interaction cloning molecular
摘 要：To investigate the interaction domains between macrophage migration inhibitory factors(MIF) and the extracellular segment of type- II trans-membrane protein CD74 using yeast two-hybrid *** using molecular cloning techniques,the DNA fragments encoding MIF,MIF and MIF were introduced into the pGBKT7 vector to construct the corresponding recombinant bait plasmids,and the DNA fragments encoding CD74,CD74,CD741 and CD74 were introduced into the pGADT7 vector to construct the corresponding recombinant activation domain(AD) plasmids,respectively. By using PEG/ LiAC method,the above 3 recombinant bait plasmids were transformed into the chemical competent yeast AHI09 cells with the 4 recombinant AD plasmids in one-one assembly manner,*** the transformed yeast AH109 cells were screened consecutively on the SD/-Trp-Leu and the SD/-Trp-Leu-Ade-His/X-a-gal nutritional *** results of restrictive endonuclease digestion and DNA sequencing proved that all the recombinant plasmids used in this study were *** yeast AHI09 cells transformed with each of the 3 recombinant bait plasmids could grow on the SD/-trp nutritional media without autonomous activation effect on the reporter gene MEL1,and the yeast AH109 cells transformed with each of the 4 recombinant AD plasmids could also grow on the SD/-leu nutritional media without autonomous activation effect on the reporter gene *** the yeast AHI09 cells co-transformed respectively with MIF,MIF,MIF expression plasmid and CD7473-232 expression plasmid could grow on the SD/-Trp-Leu-Ade-His nutritional media with the activation on the transcription of reporter gene *** interacts with the integrity extracellular segment of CD74(CD74) independent of the function domain of MIF.To investigate the interaction domains between macrophage migration inhibitory factors(MIF) and the extracellular segment of type- II trans-membrane protein CD74 using yeast two-hybrid *** using molecular cloning techniqu