ABSTRACT
OBJECTIVE: To construct Ag85A-HA2 prokaryotic expression vector, express the fusion protein and study the immunity efficacy of fusion protein against influenza A virus. METHODS: Ag85A-HA2 prokaryotic expression vector was constructed and induced with IPTG. The fusion protein was identified by SDS-PAGE and purified with His-Tag affinity chromatography. The BALB/c mice were immunized with fusion protein. Then the pathological section, lung index, lung inhibitory rate and death-protection rate were tested to evaluate the immunity efficacy of fusion protein. RESULTS: pET-32a(+)/Ag85A-HA2 prokaryotic expression vector was constructed successfully. And SDS-PAGE indicated that fusion protein was expressed correctly with a molecular mass of 70 x 10(3). The lung index and death-protection rate in experimental group were 39.30% and 80%, higher than that of control group. The pathological section also demonstrated that Ag85A-HA2 fusion protein had a protective effect on murine lungs. CONCLUSION: Ag85A-HA2 prokaryotic expression vector was successfully constructed, inducible expression and the fusion protein had an immunity efficacy against influenza A virus in animal experiment.
Subject(s)
Influenza A virus , Influenza Vaccines/immunology , Orthomyxoviridae Infections/prevention & control , Recombinant Fusion Proteins/immunology , Acyltransferases/immunology , Animals , Antigens, Bacterial/immunology , Electrophoresis, Polyacrylamide Gel , Genetic Vectors , Hemagglutinin Glycoproteins, Influenza Virus/immunology , Mice , Mice, Inbred BALB CABSTRACT
OBJECTIVE: To study the inhibition of berberine (BBR) against ECV-304 apoptosis induced by Staphylococcus aureus (S. aureus). METHODS: ECV-304 cells were pre-treated with 128 microg/mL BBR for 2 h and then S. aureus was added (1:100). The viability of cells was detected by MTT (3-4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay. The morphological changes were observed by Hoechst 33258 staining. The protection of BBR for infected cells was detected by DNA Ladder. RESULTS: ECV-304 cells' viability were not obviously affected by berberine. But S. aureus induced ECV-304 cells' viability could be significantly inhibited by pre-treatment of BBR (P < 0.05). Besides S. aureus-induced ECV-304 apoptosis could be reduced, with significantly lessened apoptotic body and unobvious DNA degradation. CONCLUSION: BBR could significantly inhibit S. aureus induced ECV-304 apoptosis.
