| Summary: | 碩士 === 國立中興大學 === 園藝學系 === 82 === This project includes construction of MT binding protein
genes from Geaniue pig (MTⅡ), antifreezing protein genes
from winter floumder(AF), and insecticidal protein genes
seperated from Bacillus thuringiensis (Bt) with vector carrying
with rubisco small subumit (rbcS) and alcohol dehydrogenase
(adh) promoter sites respectively. Constructed DNAs were
transferred into hypocotyls and cotyledons of three Brassica
vegetables on agrobacterium-mediated base. We have focused on
establishing gene transter and regeneration systems on
Broccoli, Cauliflower and Pak-choi, comparisons in gene
expression led by different promoters. Also, assessments on
the possibilities of vegetables that are resistant to
Cadmium, freezing, and Plutella xylostella are made during
discussion. Results have shown that the regeneration rate of
five transgenic species among three Brassica vegetables ranges
around 1-3%, more than 95% of these plants are GUS positive.
Transferred genes are detected by southern assay among
PCR selected plantlets; transcriptions of the three
genes in transgenic plantlets are also confirmed resulting
from Northern assay. Plantlets with rbcS promoter perform a
higher RNA expression than those with CaMV 35S
promoter, no correlations are fond with different genes
and their RNA expression. Beside, results also have show that
plantlets with rbcS promoter get higher mortalities of
Plutella xylostella rather than those with CaMV 35S
promoter which are evidenced by Bt bioassay. Regenerations
of three Brassica vegetables with three distinct genes
have been observed nowadays, among which the Bt transgenic
plantlets have complete bioassays,the other AF and MTⅡ
transgenic plants have also exhibit transgenic mRNA that
need further bioassays.
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