产气荚膜梭菌α毒素C末端的三拷贝串联表达与免疫原性分析

doi:10.16431/j.cnki.1671-7236.2018.11.031

Abstract

Abstract:

This study was aimed to obtain a tandem fusion protein of three copies of CPA C-terminal (CPA_C),and evaluate its immunogenicity.Based on the known CPA sequence of Clostridium perfringens type A(GenBank accession No.:AY823400.1),CPA_C gene was optimized,three copies of CPA_C genes (GCPA_C3) were tandemly linked in the same direction with the flexible amino acid linker (GGGS) gene and synthesized.In roder to get the recombinant protein of three copies of CPA_C(rGCPA_C3),GCPA_C3 was cloned into prokaryotic expression vector pET-30a(+) for expression and purification.Reactivity of rGCPA_C3 with antiserum of Clostridium perfringens type A was detected by Western blotting.Four rabbits were immunized with rGCPA_C3 emulsified with oil adjuvant of ISA 201 to prepare antiserum and detect the neutralizing titer.The results showed that rGCPA_C3was presented predominantly in an insoluble form (inclusion bodies),and it could react with the antiserum of Clostridium perfringens type A.After the first immunization,sera from rabbits immunized with rGCPA_C3could neutralize 30 to 50 MLD/mL Clostridium perfringens type A toxin,and 70 to 100 MLD/mL after twice immunization.Moreover,rabbits in rGCPA_C3immunized group fully survived at the dose of 1 rabbit MLD of Clostridium perfringens type A toxin challenge,whereas all of the rabbits died (100%,4/4) in control groups.These data suggested that rGCPA_C3was a potential vaccine candidate for genetic engineering subunit vaccine of Clostridium perfringens type A.

Key words: Clostridium perfringens α toxin; C-terminal; recombinant expression; antigenicity

CLC Number:

S852.61⁺6.3

DU Jige, LIU Ying, ZHU Zhen, XUE Qi, ZHANG Xiukun, LI Qihong, YIN Chunsheng, KANG Kai, YAO Wensheng, CHEN Xiaoyun. Three Copies Recombinant Expression and Immunogenicity Analysis of C-terminal of Clostridium perfringens α Toxin[J]. , 2018, 45(11): 3237-3245.

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Three Copies Recombinant Expression and Immunogenicity Analysis of C-terminal of Clostridium perfringens α Toxin

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