Taura syndrome of penaeid shrimp: Cloning of viral genome fragments and development of specific gene probes

Jocelyne Mari, Jean Robert Bonami, Donald V Lightner

Research output: Contribution to journalArticle

50 Citations (Scopus)

Abstract

The ssRNA genome extracted from purified Taura Syndrome Virus (TSV) was transcribed into double-stranded, blunt-ended cDNA and was used to construct cDNA libraries either in pUC 18 or in pBluescript II KS-vectors. Twelve recombinant plasmids chosen after screening of the libraries were subjected to restriction enzyme digestions for determination of size inserts and restriction maps. Two of them, pP15 and pQ1, were selected for probe construction. The inserts, 1500 and 1300 base pairs (bp) respectively, were DIG-11dUTP-labelled and the corresponding probes were named P15 and Q1. On northern blots and dot blots, using different denaturation methods, the 2 probes hybridized specifically with extracted RNA-TSV genome, TSV and infected TS shrimp homogenates. No positive hybridization was obtained with other shrimp viruses tested [Infectious Hypodermal and Hematopoietic Necrosis Virus (IHHNV) and Hepatopancreatic Parvovirus (HPV)]. The specificity of the 2 probes was confirmed by in situ hybridization on histological sections of TS diseased shrimps.

Original languageEnglish (US)
Pages (from-to)11-17
Number of pages7
JournalDiseases of Aquatic Organisms
Volume33
Issue number1
StatePublished - May 14 1998

Fingerprint

Taura syndrome virus
Penaeidae
molecular cloning
virus
shrimp
genome
probe
gene
Infectious hematopoietic necrosis virus
Protoparvovirus
genes
denaturation
cDNA libraries
Northern blotting
in situ hybridization
plasmids
hybridization
digestion
RNA
screening

Keywords

  • Cloning
  • Dot blot hybridization
  • In situ hybridization
  • Penaeid shrimp
  • Picomavirus
  • TSV

ASJC Scopus subject areas

  • Aquatic Science
  • Ecology

Cite this

Taura syndrome of penaeid shrimp : Cloning of viral genome fragments and development of specific gene probes. / Mari, Jocelyne; Bonami, Jean Robert; Lightner, Donald V.

In: Diseases of Aquatic Organisms, Vol. 33, No. 1, 14.05.1998, p. 11-17.

Research output: Contribution to journalArticle

@article{95e076e34df54e9589ba23142e278cc6,
title = "Taura syndrome of penaeid shrimp: Cloning of viral genome fragments and development of specific gene probes",
abstract = "The ssRNA genome extracted from purified Taura Syndrome Virus (TSV) was transcribed into double-stranded, blunt-ended cDNA and was used to construct cDNA libraries either in pUC 18 or in pBluescript II KS-vectors. Twelve recombinant plasmids chosen after screening of the libraries were subjected to restriction enzyme digestions for determination of size inserts and restriction maps. Two of them, pP15 and pQ1, were selected for probe construction. The inserts, 1500 and 1300 base pairs (bp) respectively, were DIG-11dUTP-labelled and the corresponding probes were named P15 and Q1. On northern blots and dot blots, using different denaturation methods, the 2 probes hybridized specifically with extracted RNA-TSV genome, TSV and infected TS shrimp homogenates. No positive hybridization was obtained with other shrimp viruses tested [Infectious Hypodermal and Hematopoietic Necrosis Virus (IHHNV) and Hepatopancreatic Parvovirus (HPV)]. The specificity of the 2 probes was confirmed by in situ hybridization on histological sections of TS diseased shrimps.",
keywords = "Cloning, Dot blot hybridization, In situ hybridization, Penaeid shrimp, Picomavirus, TSV",
author = "Jocelyne Mari and Bonami, {Jean Robert} and Lightner, {Donald V}",
year = "1998",
month = "5",
day = "14",
language = "English (US)",
volume = "33",
pages = "11--17",
journal = "Diseases of Aquatic Organisms",
issn = "0177-5103",
publisher = "Inter-Research",
number = "1",

}

TY - JOUR

T1 - Taura syndrome of penaeid shrimp

T2 - Cloning of viral genome fragments and development of specific gene probes

AU - Mari, Jocelyne

AU - Bonami, Jean Robert

AU - Lightner, Donald V

PY - 1998/5/14

Y1 - 1998/5/14

N2 - The ssRNA genome extracted from purified Taura Syndrome Virus (TSV) was transcribed into double-stranded, blunt-ended cDNA and was used to construct cDNA libraries either in pUC 18 or in pBluescript II KS-vectors. Twelve recombinant plasmids chosen after screening of the libraries were subjected to restriction enzyme digestions for determination of size inserts and restriction maps. Two of them, pP15 and pQ1, were selected for probe construction. The inserts, 1500 and 1300 base pairs (bp) respectively, were DIG-11dUTP-labelled and the corresponding probes were named P15 and Q1. On northern blots and dot blots, using different denaturation methods, the 2 probes hybridized specifically with extracted RNA-TSV genome, TSV and infected TS shrimp homogenates. No positive hybridization was obtained with other shrimp viruses tested [Infectious Hypodermal and Hematopoietic Necrosis Virus (IHHNV) and Hepatopancreatic Parvovirus (HPV)]. The specificity of the 2 probes was confirmed by in situ hybridization on histological sections of TS diseased shrimps.

AB - The ssRNA genome extracted from purified Taura Syndrome Virus (TSV) was transcribed into double-stranded, blunt-ended cDNA and was used to construct cDNA libraries either in pUC 18 or in pBluescript II KS-vectors. Twelve recombinant plasmids chosen after screening of the libraries were subjected to restriction enzyme digestions for determination of size inserts and restriction maps. Two of them, pP15 and pQ1, were selected for probe construction. The inserts, 1500 and 1300 base pairs (bp) respectively, were DIG-11dUTP-labelled and the corresponding probes were named P15 and Q1. On northern blots and dot blots, using different denaturation methods, the 2 probes hybridized specifically with extracted RNA-TSV genome, TSV and infected TS shrimp homogenates. No positive hybridization was obtained with other shrimp viruses tested [Infectious Hypodermal and Hematopoietic Necrosis Virus (IHHNV) and Hepatopancreatic Parvovirus (HPV)]. The specificity of the 2 probes was confirmed by in situ hybridization on histological sections of TS diseased shrimps.

KW - Cloning

KW - Dot blot hybridization

KW - In situ hybridization

KW - Penaeid shrimp

KW - Picomavirus

KW - TSV

UR - http://www.scopus.com/inward/record.url?scp=0032516224&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0032516224&partnerID=8YFLogxK

M3 - Article

C2 - 9653455

AN - SCOPUS:0032516224

VL - 33

SP - 11

EP - 17

JO - Diseases of Aquatic Organisms

JF - Diseases of Aquatic Organisms

SN - 0177-5103

IS - 1

ER -