Retinol\|binding protein(RBP)is the transport protein of vitamin A and is one of the members in the family of proteins which may all bind small hydrophobic molecules.In order to study the structure\|function relations...Retinol\|binding protein(RBP)is the transport protein of vitamin A and is one of the members in the family of proteins which may all bind small hydrophobic molecules.In order to study the structure\|function relationship of RBP,the plasmid pET3a\|RBP was constructed,and RBP was expressed in E.coli Bl21(DE3)successfully.The expressed protein,with molecular weight of about 21 kD,was about 36.95 percent of the total bacteria protein as detected by SDS\|PAGE and densitometry analysis.Then the production was confirmed by probing Western blot of E.coli lysate separated on SDS\|PAGE with antisera against hRBP.In succession,the fluorescence enhancement properties and shift in absorption spectrum had also confirmed the production’s functional characteristics.It could bind ROH in ratio of 1∶1 and K d=1.5×10 -7 mol/L.展开更多
文摘Retinol\|binding protein(RBP)is the transport protein of vitamin A and is one of the members in the family of proteins which may all bind small hydrophobic molecules.In order to study the structure\|function relationship of RBP,the plasmid pET3a\|RBP was constructed,and RBP was expressed in E.coli Bl21(DE3)successfully.The expressed protein,with molecular weight of about 21 kD,was about 36.95 percent of the total bacteria protein as detected by SDS\|PAGE and densitometry analysis.Then the production was confirmed by probing Western blot of E.coli lysate separated on SDS\|PAGE with antisera against hRBP.In succession,the fluorescence enhancement properties and shift in absorption spectrum had also confirmed the production’s functional characteristics.It could bind ROH in ratio of 1∶1 and K d=1.5×10 -7 mol/L.
