鼠源CD40L的原核表达及其对河豚毒素半抗原免疫增强效果分析

doi:10.16431/j.cnki.1671-7236.2019.08.030

Abstract

Abstract:

The aim of this study was to express recombinant protein CD40L from mice by E.coli expression system, and investigate its immunopotentiating effect on tetrodotoxin (TTX) artificial complete antigen in BALB/c mice.Total RNA from spleen of BALB/c mice was extracted by Trizol reagent and retranscribed into cDNA.Primers were designed according to CD40L CDS region, and the target gene was amplified by PCR.The recombinant vector pGEX4T-1 was constructed for prokaryotic expression, and the recombinant CD40L protein was purified.TTX-BSA and TTX-OVA were prepared by formaldehyde method according to Mannich reaction principle.The adjuvant of artificial recombinant protein CD40L was used as the experimental group and the adjuvant of Freund's was used as the control group.BALB/c mice were immunized.The immune effects of each group was analyzed using ELISA method combined with SPSS 19.0 software to explore the effect of recombinant CD40L protein on the immune effect during TTX-BSA immunization.This experiment successfully amplified 783 bp CD40L gene, prokaryotic expression and purification of GST-tagged 55 ku mice CD40L recombinant protein.Compared with TTX-BSA synergistic immunization in mice, CD40L had extremely significant immune enhancement effect at the initial stage of immunization (P<0.01) than Freund's adjuvant.In conclusion, the recombinant protein CD40L and TTX-BSA complete antigen synergistically immunized mice could enhance the body's response to hapten at the initial stage of immunization, which laid a foundation for further development of immunopotentiation adjuvants suitable for the efficient preparation of small hapten antibodies.

Key words: CD40L; adjuvant; tetraodotoxin; immune effect

CLC Number:

S852.4

PAN Yue, CAO Qi, REN Honglin, HU Pan, LI Yansong, ZHOU Yu, LIU Zengshan, LU Shiying. Prokaryotic Expression of Murine CD40L and Its Immunopotentiating Effect on Tetrodotoxin Hapten[J]. , 2019, 46(8): 2438-2445.

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Prokaryotic Expression of Murine CD40L and Its Immunopotentiating Effect on Tetrodotoxin Hapten

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