牛病毒性腹泻病毒E2基因在大豆叶片中的表达及对小鼠免疫指标的影响OA

This study aimed to investigate the expression of the bovine viral diarrhea virus(BVDV)E2 gene in soybean leaves and the immunogenicity of its expressed product.First,the neutralizing epitope sequence of the E2 gene from a BVDV Jilin isolate was analyzed,and a multi-epitope polymer gene sequence named BVDV-JZ was designed and synthesized.Subsequently,the recombinant virus SMV3-2-BVDV-JZ was successfully constructed using homologous recombination technology.Reverse transcription-polymerase chain reaction(RT-PCR),enzyme-linked immunosorbent assay(ELISA),and Western blot analysis confirmed that the BVDV-JZ multi-epitope protein was successfully expressed in transgenic soybean leaves and could be specifically recognized by BVDV-positive serum.To evaluate its oral immunization effect,BALB/c mice were immunized by gavage with 10 mg of total protein extracted from soybean leaves containing the target protein.The results showed that the BVDV-JZ multi-epitope protein could be expressed in soybean leaves.Western blot detection indicated that the BVDV-JZ protein expressed in soybean leaves was specifically recognized by BVDV-positive serum.After oral immunization in BALB/c mice,no adverse effects on growth were observed.The levels of immunoglobulin G(IgG)in serum and immunoglobulin A(IgA)in feces were significantly higher than those in the blank group(P<0.05).After 21 days of gavage,the serum levels of interferon-γ(IFN-γ)and interleukin-4(IL-4)were significantly higher than those in the blank group(P<0.05).Moreover,lymphocyte proliferation ability was enhanced after oral immunization.This study shows that soybean can serve as a bioreactor for producing exogenous proteins and can induce immune responses in vivo,providing a reference for the development and application of edible vaccines.