Taura syndrome virus (TSV), the epidemic agent of Taura syndrome (TS), was tentatively classified in the family Dicistroviridae, caused disastrous losses with high mortality rates from 60 to >90% of affected pond-cultured Litopenaeus vannamei. The amino acid sequence alignments of ORF1 revealed sequence motifs characteristic of a helicase, a protease and an RNA-dependent RNA polymerase, similar to the non-structural proteins of picornavirus, suggesting perhaps a similar to picornavirus polyprotein processing. Proteolytic processing of the TSV polyprotein and protease is still unclear. In order to further understand the role of protease interaction with polyprotein, several clones were constructed. In this study, we constructed several clones that expressed the TSV virus coat protein(VP-polyprotein)¡]pTSVVP¡^, putative 3C protease¡]pTSV3C¡^, protease¡ÏRdRp-polyprotein(pET3CD) and both VP+3C proteins(pTSVVP3C) in E. coli. The VP-polyprotein band was excised from SDS-polyacrylamide gels and electroeluted for antibody production in mice. Processing of the VP-polyprotein by 3C protease was carried out in the in vitro co-transcription/translation system and detected by Western blot. The results showed there were protein bands corresponding to the sizes of VP2+VP1 and VP1+VP3 indicating that the processing might have been partial or incomplete.
| Identifer | oai:union.ndltd.org:NSYSU/oai:NSYSU:etd-0909105-105653 |
| Date | 09 September 2005 |
| Creators | Hsiu, Lu-Chieh |
| Contributors | Liu-Jong Kang, Lin-Chan-Shing, Hsu-Chi Hsin |
| Publisher | NSYSU |
| Source Sets | NSYSU Electronic Thesis and Dissertation Archive |
| Language | Cholon |
| Detected Language | English |
| Type | text |
| Format | application/pdf |
| Source | http://etd.lib.nsysu.edu.tw/ETD-db/ETD-search/view_etd?URN=etd-0909105-105653 |
| Rights | withheld, Copyright information available at source archive |
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