Expression of GFP Gene in Sperm, Eggs, and Larvae in Koi Fish Transgenesis Using the SMGT Method with cβa-GFP Promoter

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This study aimed to determine the effectiveness of homologous β-actin promoter concentration from the common carp in expressing the GFP (Green Fluorescent Protein) marker gene during the transgenesis process of koi fish using electroporation and SMGT (Sperm Mediated Gene Transfer) methods on sperm, embryos, and larvae of Koi fish. The research employed an experimental method using a completely randomized design (CRD), with three concentrations of cβa-GFP: 10, 30, and 50ng/ μl.The electroporation treatment was conducted using an electric voltage of 30 volts and four pulses. The observed parameters include the fluorescence and integration region of cβa-GFP in sperm, eggs, and larvae, as well as the identification ofGFP in embryos and larvae, and sperm quality. The results showed that the use of the cβa (Cyprinus carpio β-actin) promoter successfully expressed GFP marker gene fluorescence in sperm, embryos, and larvae of koi fish, with the best concentration being 10ng/ μl.

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