Expression Patterns Of Hsp90 On Temperature Treatment And Vibrio Anguillarum Infection In Paralichthys Olivaceus

Paralichthys olivaceus is one of an important marine cultured fish and the sustainable development of its culture easily affected by frequently occurred diseases. The heat shock protein (HSPs) is a kind of chaperonins which has an increased expression after affection with high temperature and other adversity stimulation. Understanding the mechanism of P.olivaceus's HSPs'action is crucial to its disease prevention. In this study, the expression variation of Hsp90a and Hsp90? was studied under thermal stimulation and adventitious infection. It provides theoretical guidance for the disease control of P. olivaceus. The contents and results of this study are mainly as follows:Research 1:The expression distribution of Hsp90 gene in different developmental stages and body tissues. The real-time fluorescent quantitative PCR method was used to detect the Hsp90a and Hsp90? gene mRNA expression in different developmental stages of P. olivaceus. The results showed that Hsp90a and Hsp90? were expressed in Polivaceus's organization at different developmental stages (liver, spleen, head-kidney, metanephros, heart, muscle, skin, stomach, intestine, gill, pelvic fin, brain and blood). The Hsp90? expression level is higher than the Hsp90a in general. In liver, gill, back skin, back muscle, pelvic fin, the Hsp90? expression at 8-month P. olivaceus of age is significantly higher than other development period (P<0.05), while the Hsp90P expression quantity in different size P. olivaceus showed obvious differences.Research 2:The Hsp90 gene expression variant with thermal stimulation and vibrio anguillarum infection. The 8 months of age P. olivaceus were chosen for study, after 1 hr thermal stimulation in 5-32?, the Hsp90a expression level in spleen and liver showed dramatically elevation in the high temperature (22-32?) whereas the expression level of Hsp90? showed not obviously change. After continuous stimulation 8 hr at 10? and 28? respectively, Hsp90? and Hsp9O? of mRNA expression level showed the former low-high trend. The range ability of Hsp90? expression was significantly higher than that of changes in Hsp90? during high temperature. After 72 hr vibrio anguillarum infection, Hsp90? and Hsp90? gene expression level in P. olivaceus's spleen decreased first then increased, the Hsp90? rose significantly (P<0.05). The results showed that Hsp90? was sensitive to the hyperthermia stress and Hsp90? was extremely sensitive to the pathogen infection stimulation.Research 3:Study on P. olivaceus Hsp90? prokaryotic protein expression. The Hsp90? cDNA was inserted to the plasmid vector pBAD/g?A. The recombination vector plasmid named as pBAD/g?A-Hsp90 transferred into E.coli top 10 competent cells, induced with L-Arab sugar. The target protein was identified with SDS-PAGE electrophoresis and mass spectrum analyzing. The results showed that the recombinant proteins expressed well and this provided foundation research on follow-up Hsp90a function study.