National Taiwan University

National Science Council

Other

Chung-Hsiung Wang

Chu-Fang Lo

Chih-Yu Wu

Guang-Hsiung Kou

Chang-Jen Huang

Chih-Ming Chou

Shih-Chia Yeh

Guang-Hsung Kou

Song-Tay Lee

The present invention relates tocell lines established from pupal tissues ofincluding NTU-MV, two subclones of NTU-MV: NTU-MV-1 and NTU-MV-2. The NTU-MV cell line was confirmed to be derived fromand NTU-MV-1, and NTU-MV-2 were confirmed to be derived from NTU-MV by karyotype analysis, Internal transcribed spacer (ITS) analysis and isozyme analysis. The genotypes and characteristics of the abovementioned 3 cell lines are totally different from other insect cell lines so that they are newly established cell lines. In addition, these three MV cell lines are susceptible tomultiple nucleopolyhedrovirus (MaviMNPV). Therefore the 3 cell lines according to invention can be applied in multiplication of MaviMNPV in vitro to produce biopesticides in pest control. In addition, the cell lines can also be used as hosts for the expression vectors of baculovirus to produce recombinant proteins.

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The present invention relates tocell lines established from pupal tissues ofSwinhoe, including NTU-LY-1, NTU-LY-2, NTU-LY-3, and NTU-LY-4. These four cell lines were confirmed to be newly established cell lines derived fromby random amplified polymorphic DNA-polymerase chain reaction (RAPD-PCR) and isozyme analysis. The genotypes and characteristics of the abovementioned cell lines are totally different from other insect cell lines. In addition, these fourcell lines are susceptible to insect baculovirus ofmultiple nucleopolyhedrovirus (LyxyMNPV), LdMNPV-like virus, PenuMNPV, as well as microsporidia ofsp. (isolated from) andand the like. Therefore the invention can be applied in multiplication of the abovementioned insect-pathogenic microorganisms to produce biopesticides in pest control. In addition, the cell lines can also be used as hosts for the expression vectors of said baculovirus to produce recombinant proteins.

This invention relates to the identification, purification and detection of a new infectious viral agent in arthropods, especially shrimps. The virus is named as WSBV (Baculovirus associated with white spot syndrome). Two WSBV genomic DNA libraries were constructed and based upon the sequence of one of the cloned WSBV DNA fragments, a WSBV specific primer set for PCR to detect the WSBV infection in penaeid shrimps has been developed. The results of the present invention provide an effective diagnostic tool for screening WSBV infection in animal host organisms, in particular shrimps, to prevent the further spread of this viral disease.

Identification, purification and detection of WSBV baculovirus associated with white spot syndrome

Identification, purification and detection of WSBV (Baculovirus

Two pairs of PCR primer mixtures to be used to detect baculovirus infection: (i) a mixture of 25 to 35-base oligonucleotides including, respectively, 5'-TTTTGACGCAAATYYTAGACGCCGT-3' (SEQ ID NO: 1), and a mixture of 19 to 29-base oligonucleotides, including, respectively, 5'-TCARYATKGATTGAATWTC-3' (SEQ ID NO: 2); and (ii) a mixture of 30 to 40-base oligonucleotides, including, respectively, 5'-TAYGTGTACGACAACAARTAY-3' (SEQ ID NO: 3), and a mixture of 30 to 40-base oligonucleotides, including, respectively, 5'-GCGTCKGGYGCAAAYTCYTTWACY-3' (SEQ ID NO: 7).

Oligonucleotides for detection of baculovirus infection

Growing community of inventors

Taipei, Taiwan

Chung-Hsiung Wang