• Journal of Aquaculture
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• v.10 no.1
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• pp.33-37
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• 1997

construct containing reporter gene(pFV4CAT) regulated by carp $\beta$-actin promoter was microinjected into the one-cell stage egg of mud loach (Misgurnus mizolepis), and was successfully expressed, possibly by the integration into the genome. Both mean hatching success and early survival of the microinjected groups were not significantly different with those of control groups (P>0.05). The incidence of transgene was ranged from 7 to 48% based on the PCR and/or Southern blot analyses with the DNA prepared from fin or blood tissue. The spatial and temporal patterns of expression of the pFV4CAT gene, measured by in situ immunohistochemical analysis peroxidase-conjugated anti-CAT antibody, were variable among the experimental individuals. These results suggest that carp $\beta$-actin promoter is effective to express other transgene in mud loach, such that this promoter can be useful in the generation of valuable transgenic mud loach.

• Journal of Aquaculture
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• v.10 no.4
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• pp.409-415
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• 1997

The carp $\beta$-actin regulatory sequences and RSV/LTR promoter were tested whether they are functinal to express linked structure gene (chloramphenicol acetyltransferas, CAT) in olive flounder (Paralichthys olivaceus) by determining the patterns of gene expression following intramuscular in vivo direct injection. The injection experiments with various concentrations of both pRSVCAT and pFV4CAT clearly revealed the effectiveness of DNA dosage on expression of CAT. The increase of CAT activity was linear in both plasmids, and maximal CAT activity was obtained with 100 ug of pFV4CAT injection. The amounts of CAT expression with pFV4CAT-injected fist were higher than those with pRSVCAT-injected fish. CAT activity was readily detectable as early as one day after injection, slightly increased at day 2, and declined over time. Most amount of DNA intramuscularly injected into olive flounder muscles persisted extrachromosomally without showing any integrated or replicated form in vivo.

• Korean Journal of Ichthyology
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• v.9 no.1
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• pp.91-98
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• 1997

The transgene, pFV4CAT, containing CAT reporter gene regulated by carp $\beta$-actin promoter, was expressed in independent transgenic mud loach germ lines, determined by reverse transcriptase-PCR (RT-PCR) and enzyme-linked immunosorbant assay (ELISA). Expression of the transmitted transgene was found to be tissue-specific in F1 and F2 generations. Tissue specificity of the expression was dependent on each transgenic line with reproducible patterns. Liver and spleen did express the transgene more frequently than other tissues tested, and muscle and heart revealed the higher amount of CAT than other tissues, while testes showed the lowest expression level. The highest level of CAT expression in muscle from a transgenic F1 line was corresponding to 68-fold compared to the basal levels of controls.