Expression and transmission of transgenes in tilapia (Oreochromis niloticus)
Expression and transmission of transgenes in tilapia (Oreochromis niloticus)
The efficiency of expression of three lacZ reporter genes under the transcriptional control of the carp and the rat β-actin promoter were compared in transgenic tilapia. The gene construct containing the 4.6kb carp β-actin promoter was found to be more efficient at driving expression of the lacZ reporter gene in developing tilapia embryos compared to those containing the 4.6kb rat β-actin promoter and the 2.6kb carp β-actin promoter. The levels of expression were variable among and within the groups of embryos analyzed. The β-Galactosidase activity of the embryos was assayed both by in situ staining using X-Gal as a substrate and by a fluorometer on homogenates using Methyl umbelliferyl β-D Galactoside as substrate. The mean incidence of expression of the gene construct containing the 4.6kb carp β-actin promoter was 64.8% while that of the gene construct containing the 4.6kb rat and the 2.6kb carp β-actin promoter were 27.8% and 17.4% respectively. Variation in the level of expression was also observed. The mean fluorometer reading for the β-Galactosidase activity were 11060, 1611, and 3213 for the gene construct containing the 4.6kb carp, 4.6kb rat and the 2.6kb carp β-actin promoter respectively. It was concluded that the gene construct with regulatory sequences from the carp expresses more efficiently in tilapia when compared to the equivalent construct from the rat. In addition it was also observed that the longer carp promoter enhances expression several fold over the short carp promoter. The promoterless construct containing the lacZ reporter gene gave a very low incidence of possible positives.
University of Southampton
1995
Alam, Md. Samsul
(1995)
Expression and transmission of transgenes in tilapia (Oreochromis niloticus).
University of Southampton, Doctoral Thesis.
Record type:
Thesis
(Doctoral)
Abstract
The efficiency of expression of three lacZ reporter genes under the transcriptional control of the carp and the rat β-actin promoter were compared in transgenic tilapia. The gene construct containing the 4.6kb carp β-actin promoter was found to be more efficient at driving expression of the lacZ reporter gene in developing tilapia embryos compared to those containing the 4.6kb rat β-actin promoter and the 2.6kb carp β-actin promoter. The levels of expression were variable among and within the groups of embryos analyzed. The β-Galactosidase activity of the embryos was assayed both by in situ staining using X-Gal as a substrate and by a fluorometer on homogenates using Methyl umbelliferyl β-D Galactoside as substrate. The mean incidence of expression of the gene construct containing the 4.6kb carp β-actin promoter was 64.8% while that of the gene construct containing the 4.6kb rat and the 2.6kb carp β-actin promoter were 27.8% and 17.4% respectively. Variation in the level of expression was also observed. The mean fluorometer reading for the β-Galactosidase activity were 11060, 1611, and 3213 for the gene construct containing the 4.6kb carp, 4.6kb rat and the 2.6kb carp β-actin promoter respectively. It was concluded that the gene construct with regulatory sequences from the carp expresses more efficiently in tilapia when compared to the equivalent construct from the rat. In addition it was also observed that the longer carp promoter enhances expression several fold over the short carp promoter. The promoterless construct containing the lacZ reporter gene gave a very low incidence of possible positives.
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Published date: 1995
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Local EPrints ID: 458659
URI: http://eprints.soton.ac.uk/id/eprint/458659
PURE UUID: d2741777-260c-4a8a-95f1-140a3ebc22b3
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Date deposited: 04 Jul 2022 16:53
Last modified: 04 Jul 2022 16:53
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Author:
Md. Samsul Alam
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