TAURA SYNDROME OF PENAEID SHRIMP - CLONING OF VIRAL GENOME FRAGMENTS AND DEVELOPMENT OF SPECIFIC GENE PROBES

The ssRNA genome extracted from purified Taura Syndrome Virus (TSV) wa s transcribed into double-stranded, blunt-ended cDNA and was used to c onstruct cDNA libraries either in pUC 18 or in pBluescript II KS-vecto rs. Twelve recombinant plasmids chosen after screening of the librarie s were subjected to restriction enzyme digestions for determination of size inserts and restriction maps. Two of them, pP15 and pQ1, were se lected for probe construction. The inserts, 1500 and 1300 base pairs ( bp) respectively, were DIG-11dUTP-labelled and the corresponding probe s were named P15 and Q1. On northern blots and dot blots, using differ ent denaturation methods, the 2 probes hybridized specifically with ex tracted RNA-TSV genome, TSV and infected TS shrimp homogenates. No pos itive hybridization was obtained with other shrimp viruses tested [Inf ectious Hypodermal and Hematopoietic Necrosis Virus (IHHNV) and Hepato pancreatic Parvovirus (HPV)]. The specificity of the 2 probes was con firmed by in situ hybridization on histological sections of TS disease d shrimps.