Two-Component And ABC-Transporter Proteins Interacted By STPK And STPP Proteins In Streptococcus Suis Type Ⅱ

Streptococcus suis type2 is a kind of important zoonoses pathogen, which caused huge losses to aquaculture in our country in recent years, however, its pathogenic mechanism is not clear. In prokaryotes, Serine/threonine protein kinases(STPK) and Serine/threonine phosphatases(STPP)can make the specific substrate protein serine/threonine phosphorylation/phosphorylation and catalytic function proteins phosphorylation/phosphorylation, Which involved in the complex signaling pathway. STPK and STPP can directly or indirectly control gene transcription, metabolic pathway, immune regulation, division of bacteria and virulence and stress reaction. The ABC-transporter system of Bacteria is an important component of the cellular material transportation system. ABC-transporter system is associated with bacterial growth and virulence in streptococcus suis type 2, so we infer it may participate in regulation system of STPK protein and STPP protein. In addition, the bacterial two-component system (TCS) is widely distributed in bacteria, which can regulate the expression of organisms genetic to adapt to the change and stimulation of external environment. And TCS can regulate the cell cycle, pathogenicity, etc, which plays an important role in physiological processes, so we infer it May also be involved in regulation system of STPK protein and STPP protein.This topic mainly Identify the protein interaction networks between STPK/STPP and the system of ABC-transporters/TCS by the BacterioMatch Ⅱ two-hybrid system in streptococcus suis type 2. And we have got twenty eight groups meaningful interactions.(1) In this topic,I completed gene cloning of the ABC-transporter and two-component libraries, in ABC transporter library for a total of Two hundred and eight clonings; in two-component library for a total of fifty-two clonings.In addition, Ⅰ Assisted laboratory members establish gene library of streptococcus suis type 2.(2) I have got seven proteins which is interacted with STPP by the BacterioMatch Ⅱ two-hybrid system, And complete activated authentication of the BacterioMatch Ⅱ two-hybrid. In the ABC- transporters system, I have got six protein interactions with STPP protein. Screening to ABC-transporters proteins respectively related with phosphate, atpase, sugar, amino acid transport. In the two-component system, I got one protein interactions with STPP protein. This is a histidine kinase of two-component system.(3) I have got seven proteins which is interacted with STPK by the BacterioMatch Ⅱ two-hybrid system, And complete activated authentication of the BacterioMatch Ⅱ two-hybrid. In the ABC-transporters system, I have got sixteen protein interactions with STPK protein. Screening to ABC-transporters proteins respectively related with glutamate dehydrogenase, pentanoic acid kinase serine, threonine phosphatase, multi-drug transporters, the substrate binding protein for phosphate transport, polysaccharide, enzyme, metal ion, ATP enzyme. In the two-component system, I have five protein interactions with STPK protein. The two-component protein are histidine kinase and Reaction to the regulators.(4) Based on results of the BacterioMatch Ⅱ two-hybrid, I selected the STPK protein and four Reaction to the regulators of two-component system in streptococcus suis type 2 as the research objects. And respectively build pGEX-KG and pET-28 c protein expression plasmid. At the same time,I Carried out expression and purification of proteins, and laid a solid foundation of GST-pull down experiment for the next step.