LIN Jing-Nan, ZHAO Jing-Zhuang, LIU Miao, REN Guang-Ming, LU Tong-Yan, XU Li-Ming. ESTABLISHMENT OF YEAST EXPRESSION SYSTEM FOR SPRING VIRERNIA OF CARP VIRUS GLYCOPROTEIN[J]. ACTA HYDROBIOLOGICA SINICA, 2019, 43(5): 977-982. DOI: 10.7541/2019.116
Citation: LIN Jing-Nan, ZHAO Jing-Zhuang, LIU Miao, REN Guang-Ming, LU Tong-Yan, XU Li-Ming. ESTABLISHMENT OF YEAST EXPRESSION SYSTEM FOR SPRING VIRERNIA OF CARP VIRUS GLYCOPROTEIN[J]. ACTA HYDROBIOLOGICA SINICA, 2019, 43(5): 977-982. DOI: 10.7541/2019.116

ESTABLISHMENT OF YEAST EXPRESSION SYSTEM FOR SPRING VIRERNIA OF CARP VIRUS GLYCOPROTEIN

  • In this study, open reading frame (ORF) of glycoprotein (1530 bp) was amplified by using RNA extracted from Spring Virernia of Carp Virus (SVCV). The SVCV G ORF was cloned into pYD1 vector to construct a recombi-nant plasmid pYD1-G and then transformed into competent yeast cells EBY100, and positive colonies were screened by colony PCR. The expression of G gene was induced by 2% glucose and detected by cell immunofluorescence and flow cytometry. The immunofluorescence staining observed increased EBY100-pYD1-G signal of induced yeast cells with increased induction time. Flow cytometry analysis observed significantly increased fluorescence intensities in prolonged induced EBY100-pYD1-G cells (P<0.05). These results indicated the SVCV G protein has been successfully expressed and localized on the surface of yeast cell. This study laid a foundation for the novel oral vaccine development against SVCV infection in carps in future.
  • loading

Catalog

    /

    DownLoad:  Full-Size Img  PowerPoint
    Return
    Return