Cytoplasmic Proteins And Protein Complexome Related With Salt-regulation In Escherichia Coli K12

Cytoplasmic proteins play an important role against the outside environment. The changes of the bacteria's surroundings usually take on as the changes of the osmolarity.Therefore,researches into different expression of bacterial cytoplasmic proteins under different osmolarity become very significant.It will be helpful to know about the cytoplasmic proteins' mechanism and the role of the cytoplasmic proteins in the bacteria.This paper took E.coli K-12 as the research object.First,we identified 6 cytoplasmic proteins that express differently in different osmolarity,using high-through-put and high-resolution 2-DE electrophoresis gels and high sensitive mass spectrometry tool.Then,we validated the expressive difference by using proteomic and genetic methodologies.Importantly,our further proteomic studies speculated 3 protein complexes of the salt regulative proteins by the use of native/SDS-PAGE,spectrometry tool and databank.Then,we validated the protein complexes through Western blotting, Far-Western blotting and Co-IP.At last,we tested separately the effects of the over-expression strain and the gene-knockout strain to the growth of E.coli K-12 in different salt concentration.Finally,gene engineering methodologies were applied to gain the clone and over-expression strains of some two-component system proteins.And then we obtained the anti-serum of each two-component system protein.In succession,we discussed the relationship between the two-component system proteins and the adaptability of E.coli K-12 in different salt concentration by Western blotting.The results revealed that PhoP,PhoR and RcsF may have a relativity with it.