Expression Of The Retinol-binding Protein And Interaction With Other Proteins

Retinol-binding protein (RBP) is a retinol-specific binding protein. It is a hydrophobic protein which is made of 184 amino acids and its mass is about 21KD. In the first part of the experiment the retinol- binding protein's prokaryotic expression vector was constructed. Then this vector was transferred into the E coli. to express the RBP. After detected by SDS-PAGE electrophoresis we found that the retinol binding protein expressed by the E coli. was in the form of the inclusion. As it is already known that the inclusion did not have its biological activity and it was very complicated to separate and purify protein from the inclusion. So in the second part of this experiment, we constructed the eukaryotic vector of RBP named pPIC9—RBP, then this vector was transfered into the Pichia yeast cell GS115. The positive cell was screened by SDS-PAGE electrophoresis. In the last part of this experiment, we discussed and did some research about the interaction of the RBP with other proteins. The main methods were immune precipition and Western blot. Through this experiments we could comprehend the transportation and the absorption of the vitamin A .And grab some clues about the diseases related with vitamin A.