Academia Sinica

National Taiwan Ocean University

Jen-Leih Wu

Jyh-Yih Chen

Jiann-Ruey Hong

Guor Mour Her

Chi-Yao Chang

Jenn-Kan Lu

Shao-Yang Hu

Tze-Ting Chiou

A method of inducing male and/or female infertility in a genetically modified (GM) fish is disclosed. Also disclosed is a method of generating an infertile GM fish with a phenotype and/or genotype of interest. The method involves generation of a GM fish whose genome comprises a foreign transgene operably linked to a fish gonad-specific promoter selected from the group consisting of an ovary-specific promoter and a testis-specific promoter. The transgene comprises a suicide gene selected from the group consisting of a reductase and a photosensitizer, in which the reductase is operably linked to a reporter gene. Infertility of the GM fish is induced if the GM fish expressing the reductase is treated with an effective amount of a reductase-activated cytotoxic prodrug or if the GM fish expressing the photosensitizer is treated with light irradiation.

Infertility control of genetically modified fish

The present invention provides an antimicrobial peptide, monodoncin, which is isolated and purified fromand is capable of being mass produced by molecular cloning techniques in a heterologous expression system, such as yeast. Monodoncin demonstrates a wide-range of bacteriostatic and bactericidal effects on G(&#x2212;) and G(+) bacteria as well as fungicidal activities, and can be used in combination with conventional antibiotics as 'cocktail therapy' to improve the therapeutic effects of the conventional antibiotics.

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The present invention relates to expression control sequences of a vertebrate liver fatty acid binding protein (L-FABP) gene that, when operably linked to a reporter (e.g., a heterologous reporter, such as the green fluorescent protein (GFP)), directly express the reporter in a fashion that mimics the liver-specific development of the L-FABP gene in the vertebrate.

Transgenic fish germline expression driven by liver fatty acid binding (L-FABP) gene promoter and applications thereof

The present invention provides a mechanism for studies of apoptosis in aquatic organisms by infecting the aquatic organisms with aquabirnavirus, especially infectious pancreatic necrosis virus (IPNV). The infection of IPNV in an aquatic cell such as a Chinook salmon embryo cell (CHSE-214) converts the cell into an apoptotic cell. The present invention also provides a method for monitoring the morphological changes during apoptosis by cloning EGFP (a variant type of GFP) to an aquatic cell and monitoring the fluorescence using microscopic techniques. The intensity of the fluorescence reflects the expression of EGFP in cells. Finally, the present invention provides means for inducing or preventing/rescuing apoptosis in a host, which include aquatic and vertebrate. The apoptosis can be induced by IPNV infection or VP3 transfection that VP3 is a 32 kDa protein derived from IPNV segment A. The apoptosis can be prevented or rescued by an antisense VP3 RNA or a zfMcl-1a protein.

Regulation of apoptosis in aquatic organisms by aquabirnavirus

The present invention provides a mechanism for studies of apoptosis in aquatic organisms by infecting the aquatic organisms with aquabirnavirus, especially infectious pancreatic necrosis virus (IPNV). The infection of IPNV in an aquatic cell such as a Chinook salmon embryo cell (CHSE-214) converts the cell into an apoptotic cell. The present invention also provides a method for monitoring the morphological changes during apoptosis by cloning EGFP (a variant type of GFP) to an aquatic cell and monitoring the fluorescence using microscopic techniques. The intensity of the fluorescence reflects the expression of EGEP in cells. Finally, the present invention provides means for inducing or preventing/rescuing apoptosis in a host, which include aquatic and vertebrate. The apoptosis can be induced by IPNV infection or VP3 transfection.that VP3 is a 32 kDa protein derived from IPNV segment A. The apoptosis can be prevented or rescued by an antisense VP3 RNA or a zfMcl-1a protein.

The present invention relates to the finding and construction of fish insulin-like growth factor II (IGF-II) cDNAs which can be cloned and expressed in cells. This invention also relates to the production of biologically active fish IGF-II polypeptides by a gene expression system using fish IGF-II cDNAs. The fish IGF-II cDNAs have 1977 bp which transcribe into a prepeptide (signal peptide), and B, C, A, D, E domain peptides. The fish mature IGF-II is a single polypeptide containing the NH,&mdash;B&mdash;C&mdash;A&mdash;D&mdash;COOH domains. The mature IGF-II polypeptide is 7 kDa in weight and has 70 amino acids. The fish recombinant IGF-II cDNA can be cloned and expressed in,, yeast, baculovirus, and fish cells. The isolated and purified IGF-II E domain peptide has mitogenic and anti-tumor activity.

Production of biologically active recombinant insulin-like growth factor II polypeptides

This present invention relates to the findings of the DNA sequences of fish insulin-like growth factor II (IGF-II) promoter regions and recombinant IGF-II promoters. These DNA sequences are capable of being expressed in eukaryotic cells and fish embryos of another fish species. The integration of the IGF-II promoter regions or recombinant IGF-II promoters into fish of another species results in the creation of a transgenic fish. The results of this invention illustrate that a fish IGF-II promoter not only can act as a growth factor to stimulate the growth and development of fish, but also is capable of being expressed in other eukaryotic cells such as in human cells.

Fish insulin-like growth factor II promoter

The present invention relates to the finding and construction of fish insulin-like growth factor II (IGF-II) cDNAs which can be cloned and expressed in cells. This invention also relates to the production of biologically active fish IGF-II polypeptides by a gene expression system using fish IGF-II cDNAs. The fish IGF-II cDNAs have 1977 bp which transcribe into a prepeptide (signal peptide), and B, C, A, D, E domain peptides. The fish mature IGF-II is a single polypeptide containing the NH.sub.2 -B-C-A-D-COOH domains. The mature IGF-II polypeptide is 7 kDa in weight and has 70 amino acids. The fish recombinant IGF-II CDNA can be cloned and expressed in E. coli, yeast, baculovirus, and fish cells.

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Taipei, Taiwan

Jen-Leih Wu