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Title: Fate of novel DNA in transgenic rainbow trout (Oncorhynchus mykiss)
Author: Iyengar, Arati
ISNI:       0000 0001 3587 6692
Awarding Body: University of Southampton
Current Institution: University of Southampton
Date of Award: 1993
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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.
Supervisor: Not available Sponsor: Not available
Qualification Name: Thesis (Ph.D.) Qualification Level: Doctoral
EThOS ID:  DOI: Not available