Generation of transgenic medaka Oryzias curvinotus (Nichols Pope 1927) carrying a cyan fluorescent protein gene driven by Alpha Actin promoter

Thuy VY Nguyen Hoang, . and Thanh Trung Mai Nguyen, . and Quoc Binh Nguyen, . and Kieu Hoa Nguyen Thi, . and Van Du Nguyen, . and Thach Thao Luu Thi, . and Nguyen Vu Thanh, . Generation of transgenic medaka Oryzias curvinotus (Nichols Pope 1927) carrying a cyan fluorescent protein gene driven by Alpha Actin promoter. pp. 56-62. ISSN 0116-6514

The study aimed to produce fluorescent protein transgenic medaka Oryzias curvinotus (Nichols Pope 1927) as a novel strain of ornamental fish. These fish were produced by transferring a plasmid consisting of a fluorescent reporter gene and a strong promoter into one-cell stage embryos. For this purpose myosin light chain 2 but not other promoters was mainly used. The study also evaluated the stability of the transgenic medaka germline acquiring vivid fluorescent phenotypes via the transgenesis of the cyan fluorescent protein (CFP) gene under the control of O. curvinotus skeletal alpha-actin (OCacta) promoter. The pOCacta-CFP plasmid containing a OCacta promoter and CFP reporter gene was transferred into the one-cell stage of O. curvinotus embryos by a microinjection technique. As a result 36 of 1386 microinjected O. curvinotus embryos exhibited CFP signals in their trunks. The expressed CFP signals in O. curvinotus embryos and adults were detected under a microscope using a green fluorescent protein (GFP) filter (450“490 nm wavelength) and blue LED light (400“450 nm wavelength). Five O. curvinotus founders showing clear CFP signals were selected and crossed with non-transgenic counterparts to produce subsequent generations. Among strains the frequency of germline transmission from founder to F1 was highly variable. Only two of the five founders successfully pass the transgene to the F1 generation. At present the progeny of subsequent generations is being produced and tested for the expression of CFP signals and therefore stable lines are ongoing.

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Abstract

The study aimed to produce fluorescent protein transgenic medaka Oryzias curvinotus (Nichols Pope 1927) as a novel strain of ornamental fish. These fish were produced by transferring a plasmid consisting of a fluorescent reporter gene and a strong promoter into one-cell stage embryos. For this purpose myosin light chain 2 but not other promoters was mainly used. The study also evaluated the stability of the transgenic medaka germline acquiring vivid fluorescent phenotypes via the transgenesis of the cyan fluorescent protein (CFP) gene under the control of O. curvinotus skeletal alpha-actin (OCacta) promoter. The pOCacta-CFP plasmid containing a OCacta promoter and CFP reporter gene was transferred into the one-cell stage of O. curvinotus embryos by a microinjection technique. As a result 36 of 1386 microinjected O. curvinotus embryos exhibited CFP signals in their trunks. The expressed CFP signals in O. curvinotus embryos and adults were detected under a microscope using a green fluorescent protein (GFP) filter (450“490 nm wavelength) and blue LED light (400“450 nm wavelength). Five O. curvinotus founders showing clear CFP signals were selected and crossed with non-transgenic counterparts to produce subsequent generations. Among strains the frequency of germline transmission from founder to F1 was highly variable. Only two of the five founders successfully pass the transgene to the F1 generation. At present the progeny of subsequent generations is being produced and tested for the expression of CFP signals and therefore stable lines are ongoing.

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Item Type:	Article
AGROVOC Term:	Ornamental fishes
AGROVOC Term:	Animal embryos
AGROVOC Term:	Laboratory experimentation
AGROVOC Term:	Microscopes
AGROVOC Term:	Experiments
AGROVOC Term:	analysis
AGROVOC Term:	Fish industry
AGROVOC Term:	Transgenesis
AGROVOC Term:	Transgenic animals
Depositing User:	Mr. AFANDI ABDUL MALEK
Last Modified:	24 Apr 2025 00:55
URI:	http://webagris.upm.edu.my/id/eprint/9762

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