Molecular and immunogenic analysis of Jembrana disease virus Tat

摘要: Jembrana disease is an acute and severe of Bali cattle (Bos javanicus) endemic in Indonesia that caused by a bovine lentivirus designated virus (JDV). Previous studies have demonstrated it possible to induce protective immunity against the immunisation with crude whole vaccine prepared from tissues infected cattle. This has been ameliorate clinical signs resulting exposure infection but safer amenable commercial production techniques required. JDV, like all lentiviruses, encodes transcriptional trans-activator Tat protein encoded one or both two exons tat gene. particularly essential for replication was hypothesised induction immune response JDV may effect protection infection. Investigations were therefore conducted on provide basic information would enable be further investigated as potential immunogen incorporation into vaccines control disease. Analysis transcripts obtained three strains suggested that, during disease, produced at this stage process translated first coding exon only. Nucleotide variation exon, which amino acid variations protein, evident especially between geographically different regions Indonesia. There was; however, conservation functional domains cysteine-rich, core regions, single might protect heterologous strains. Subsequent reported thesis concentrated 1 strain JDV. The cloned pGEX vector recombinant expressed Escherichia coli. Methods purification developed. Immunogenicity initially inoculation sheep developed high titred specific antibody response. Antibodies induced recognised native proteins provided valuable reagent subsequent detection vitro vivo. Aspects determined had naturally experimentally JDV, compared levels immunodominant capsid protein. antibodies detected 23 % 128 disease-endemic areas Indonesia; these cattle, evidence previous Western blot analysis. In low serum month post-infection then decreased; increased over 6-month observation period following Tatantibody soon after secreted extracellularly contrast JDV-infected apparently greater injection strong animals there conformational difference poor immunogen, too As alternative means inducing Tat, perhaps associated cell-mediated rather than response, candidate DNA insertion vector. Transfection naked plasmid mammalian cells expression maintained antigenicity. results construct merits animal attempting A method assaying trans-acting function also will application quality procedures large-scale vaccine.