巴氏杆菌糖酵解酶的免疫保护效应研究

doi:10.16431/j.cnki.1671-7236.2021.06.032

Abstract

Abstract: The purpose of this study was to evaluate the immunoprotective efficacy of all 9 members of Pasteurella glycolytic enzymes as an antigen.Firstly,the glycolytic enzyme genes of Pasteurella multocida C51-17 strain were amplified by PCR and linked to pET-28a(+) plasmid.The recombinant plasmid was transferred into Escherichia coli BL21(DE3) strain and induced to express overnight.The recombinant proteins of Pasteurella glycolytic enzymes were purified from the supernatant of the collected strain after ultrasonic crushing.After SDS-PAGE,the Pasteurella glycolytic enzymes were transferred to PVDF membranes,and the recovered sera from C51-17 strain infection was incubated.ECL staining was performed after incubating the second antibody.At last,C57BL/6 mice were immunized with glycolytic enzymes.After high level of specific antibody was produced in the immunized groups,C51-17 strain was used to challenge.Sequencing results showed that the genes of all 9 members of the Pasteurella glycolytic enzymes were successfully cloned into pET-28a(+) plasmid,and the recombinant protein was obtained after induced expression and purification.Western blotting results showed that phosophoglucose isomerase (PGI),triose phosphate isomerase (TPI),glyceraldehyde-3-phosphate dehydrogenase (GPD),phosphoglycerate kinase (PGK),and pyruvate kinase (PYK) could react with the recovered sera from Pasteurella multocida C51-17 strain.The results of immunoprotection test showed that PGI,aldolase (ALD),GPD,PGK,phosphoglycerate mutase (PGM) and enolase (ENO) had some protective effects.In conclusion,the results of this study showed that some members of the Pasteurella glycolytic enzymes had a certain degree of immune protection,which provided a new basis for the prevention and control of pasteurellosis.

Key words: pasteurellosis; Pasteurella multocida; glycolytic enzyme; protective antigen; subunit vaccine

CLC Number:

S852.61^＋2

ZHU Weifeng, CHEN Lu, WANG Fang, HU Bo, RUAN Zhiyang, WANG Gaojie. Study on Immunoprotective Efficacy of Pasteurella Glycolytic Enzymes[J]. China Animal Husbandry and Veterinary Medicine, 2021, 48(6): 2188-2195.

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Study on Immunoprotective Efficacy of Pasteurella Glycolytic Enzymes

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