| Summary: | 碩士 === 國立交通大學 === 生物科技研究所 === 87 === In this study, we report the effects of twenty different substitutions in E.coli thioredoxin on two major regions of the protein: the G33 site and G74 site where are the hydrophobic areas implicated in protein/protein interactions. There single and double mutants are produced by site-directed mutagenesis. The twenty different mutants have been tested in vivo with bacterial phages. Substitutions of glycine74 by some support phage T3/T7 growth at 37 ℃ whereas the rest substitutions prevented phage T3/T7 growth to different degrees. Substitution of glycine74 by threonine and substitution of glycine33 by alanine or histidine support phage T7 growth at 37 ℃. However, the rest mutants prevented phage T7 growth. As to f1 and M13 phages, glycine74 is more important than glycine33 when E.coli thioredoxin interacts with the protein of filamentous phage. We know little about the mechanism of thioredoxin with filamentous gene I protein. But we are sured that the interaction areas are still located on the hydrophobic surface of E.coli thioredoxin, including amino acid residues 32-35,72-76, and 91-94.
|
|---|
