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Fate of novel DNA in transgenic rainbow trout (Oncorhynchus mykiss)

Fate of novel DNA in transgenic rainbow trout (Oncorhynchus mykiss)
Fate of novel DNA in transgenic rainbow trout (Oncorhynchus mykiss)

Various aspects concerning the fate of novel DNA in rainbow trout were investigated. These included mosaicism, germ-line transmission, and the methylation status of a mouse metallothionein-1/rat growth hormone gene (mMTrGH) construct, in addition to the fate and temporal expression patterns of injected DNA sequences during development, using a carp β-actin/CAT gene (pFV1) construct.

Widespread mosaicism was observed amongst the transgenic fish, resulting in an unequal distribution of transgene copies in the cells/tissues of the fish. Germ line transmission was, however, observed in 50% of the transgenic fish containing the mMTrGH gene construct, with transmission frequencies ranging from 14-81%. The lack of transmission from the rest of the founder transgenics could also be attributed to mosaicism. The methylation studies revealed tissue-specific differences in the methylation patterns of the mMTrGH transgene, but could not be correlated with expression due to the consistent lack of expression of this transgene in our founder transgenics.

With respect to the fate of DNA during development, prolonged persistence of injected DNA sequences appeared likely, probably contributing to the mosaicism observed. Studies on the conformational influences of injected DNA revealed that linear molecules were more efficient at forming concatemers and persisting through development. They, therefore, resulted in high levels of expression, and perhaps greater rates of integration. The concatemers generated with circular injected molecules appeared to consist of head to tail arrangements of transgene copies, whereas those generated with linear injected molecules appeared to consist of random arrangements of transgene copies.

University of Southampton
Iyengar, Arati
5eee66e9-e5b3-4335-8cd0-7ba1d35a9603
Iyengar, Arati
5eee66e9-e5b3-4335-8cd0-7ba1d35a9603

Iyengar, Arati (1993) Fate of novel DNA in transgenic rainbow trout (Oncorhynchus mykiss). University of Southampton, Doctoral Thesis.

Record type: Thesis (Doctoral)

Abstract

Various aspects concerning the fate of novel DNA in rainbow trout were investigated. These included mosaicism, germ-line transmission, and the methylation status of a mouse metallothionein-1/rat growth hormone gene (mMTrGH) construct, in addition to the fate and temporal expression patterns of injected DNA sequences during development, using a carp β-actin/CAT gene (pFV1) construct.

Widespread mosaicism was observed amongst the transgenic fish, resulting in an unequal distribution of transgene copies in the cells/tissues of the fish. Germ line transmission was, however, observed in 50% of the transgenic fish containing the mMTrGH gene construct, with transmission frequencies ranging from 14-81%. The lack of transmission from the rest of the founder transgenics could also be attributed to mosaicism. The methylation studies revealed tissue-specific differences in the methylation patterns of the mMTrGH transgene, but could not be correlated with expression due to the consistent lack of expression of this transgene in our founder transgenics.

With respect to the fate of DNA during development, prolonged persistence of injected DNA sequences appeared likely, probably contributing to the mosaicism observed. Studies on the conformational influences of injected DNA revealed that linear molecules were more efficient at forming concatemers and persisting through development. They, therefore, resulted in high levels of expression, and perhaps greater rates of integration. The concatemers generated with circular injected molecules appeared to consist of head to tail arrangements of transgene copies, whereas those generated with linear injected molecules appeared to consist of random arrangements of transgene copies.

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Published date: 1993

Identifiers

Local EPrints ID: 462540
URI: http://eprints.soton.ac.uk/id/eprint/462540
PURE UUID: 42407d50-89d9-4793-bde6-1ea06dcf3277

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Date deposited: 04 Jul 2022 19:18
Last modified: 04 Jul 2022 19:18

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Contributors

Author: Arati Iyengar

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