Terminal deoxynucleotidyl transferase(Td T) has been characterized as template-independent polymerase using single-stranded DNA(ss DNA) as primers to generate random oligonucleotides. However, the extension performanc...Terminal deoxynucleotidyl transferase(Td T) has been characterized as template-independent polymerase using single-stranded DNA(ss DNA) as primers to generate random oligonucleotides. However, the extension performance of Td T to single-stranded RNA(ss RNA) is vague. By systematically comparing and contrasting the performance of Td T-catalyzed ss DNA and ss RNA extension, it is indicated that the catalytic efficiency of ss RNA as primers was about 3 times lower than ss DNA as primers. Collectively, it is believed that understanding the catalytic performance of Td T will help to design the strategy to synthesize chimeric DNA on 3-OH of ss RNA, which becomes invaluable.展开更多
African swine fever virus(ASFV)infects domestic pigs and European wild boars with strong,hemorrhagic and high mortality.The primary cellular targets of ASFV is the porcine macrophages.Up to now,no commercial vaccine o...African swine fever virus(ASFV)infects domestic pigs and European wild boars with strong,hemorrhagic and high mortality.The primary cellular targets of ASFV is the porcine macrophages.Up to now,no commercial vaccine or effective treatment available to control the disease.In this study,three recombinant Saccharomyces cerevisiae(S.cerevisiae)strains expressing fused ASFV proteins-porcine Ig heavy chains were constructed and the immunogenicity of the S.cerevisiae-vectored cocktail ASFV feeding vaccine was further evaluated.To be specific,the P30-Fcγand P54-Fcαfusion proteins displaying on surface of S.cerevisiae cells were produced by fusing the Fc fragment of porcine immunoglobulin IgG1 or IgA1 with p30 or p54 gene of ASFV respectively.The recombinant P30-Fcγand P54-Fcαfusion proteins expressed by S.cerevisiae were verified by Western blotting,flow cytometry and immunofluorescence assay.Porcine immunoglobulin Fc fragment fused P30/P54 proteins elicited P30/P54-specific antibody production and induced higher mucosal immunity in swine.The absorption and phagocytosis of recombinant S.cerevisiae strains in IPEC-J2 cells or porcine alveolar macrophage(PAM)cells were significantly enhanced,too.Here,we introduce a kind of cheap and safe oral S.cerevisiae-vectored vaccine,which could activate the specific mucosal immunity for controlling ASFV infection.展开更多
基金supported by the National Natural Science Foundation of China (NSFC, Nos. 21927814 and 21772143 to J.Y. Zhang)the National Science Foundation of Tianjin (Nos. 20YDTPJC00090, 19ZXDBSY00070 and 20YFZCSY00990 to X.Q. Gong)。
文摘Terminal deoxynucleotidyl transferase(Td T) has been characterized as template-independent polymerase using single-stranded DNA(ss DNA) as primers to generate random oligonucleotides. However, the extension performance of Td T to single-stranded RNA(ss RNA) is vague. By systematically comparing and contrasting the performance of Td T-catalyzed ss DNA and ss RNA extension, it is indicated that the catalytic efficiency of ss RNA as primers was about 3 times lower than ss DNA as primers. Collectively, it is believed that understanding the catalytic performance of Td T will help to design the strategy to synthesize chimeric DNA on 3-OH of ss RNA, which becomes invaluable.
基金supported by the National Key Research and Development Program of China (2018YFD0500500)。
文摘African swine fever virus(ASFV)infects domestic pigs and European wild boars with strong,hemorrhagic and high mortality.The primary cellular targets of ASFV is the porcine macrophages.Up to now,no commercial vaccine or effective treatment available to control the disease.In this study,three recombinant Saccharomyces cerevisiae(S.cerevisiae)strains expressing fused ASFV proteins-porcine Ig heavy chains were constructed and the immunogenicity of the S.cerevisiae-vectored cocktail ASFV feeding vaccine was further evaluated.To be specific,the P30-Fcγand P54-Fcαfusion proteins displaying on surface of S.cerevisiae cells were produced by fusing the Fc fragment of porcine immunoglobulin IgG1 or IgA1 with p30 or p54 gene of ASFV respectively.The recombinant P30-Fcγand P54-Fcαfusion proteins expressed by S.cerevisiae were verified by Western blotting,flow cytometry and immunofluorescence assay.Porcine immunoglobulin Fc fragment fused P30/P54 proteins elicited P30/P54-specific antibody production and induced higher mucosal immunity in swine.The absorption and phagocytosis of recombinant S.cerevisiae strains in IPEC-J2 cells or porcine alveolar macrophage(PAM)cells were significantly enhanced,too.Here,we introduce a kind of cheap and safe oral S.cerevisiae-vectored vaccine,which could activate the specific mucosal immunity for controlling ASFV infection.
