Please use this identifier to cite or link to this item: http://hdl.handle.net/11455/29259
標題: 鎘結合蛋白基因、抗凍蛋白基因及蘇力菌殺蟲晶體蛋白轉移至三種蕓薹屬蔬菜之研究
Studies on the Cd-binding protein, antifreeze protein and Bt- toxin genes transfer into three species of Brassica spp.
作者: 尤進欽
Yiu, Jin-Chin
關鍵字: Cd-binding protein;鎘結合蛋白基因;antifreeze protein;Bt-toxin genes;Brassica;抗凍蛋白基因;蘇力菌殺蟲晶體蛋白;蕓薹屬蔬菜;基因轉移;農桿菌
出版社: 園藝學系
摘要: 
本試驗將分離自天竺鼠的鎘結合蛋白基因(MTⅡ )、比目魚抗凍蛋白基
因(AF)及蘇力菌殺蟲晶體蛋白基因( Bt),構築到攜帶有rubisco
small subunit(rbcS)及 alcohol hydrogenase( adh )啟動子的轉殖
載體,並利用農桿菌將其轉移到青花菜(綠王及翠光)、花椰菜(秋王及
鳳山極早生)及小白菜(臺農一號)的子葉或下胚軸。本試驗的目的為建
立青花菜、花椰菜及小白菜基因轉移及植株再生系統,研究不同啟動子對
表現此三種基因的影響,並探討培育成耐鎘、抗凍及抗小菜蛾之蔬菜的可
行性。本研究的結果顯示,以農桿菌轉殖供試之三種蕓薹屬的五個品種蔬
菜之再生率在 1 - 3% 之間。檢測轉殖植株之 GUS 基因,有 95% 以上呈
正反應。以南方墨點法分析 PCR 正反應之轉殖植株,均可偵測到轉移基
因存在於轉殖植株的 DNA。北方墨點法分析之結果顯示,轉移之三種基因
在轉殖植物體內均有轉錄。無論 MT-Ⅱ、AF 或 Bt 基因以rbcS啟動子質
體的轉殖後再生植株,其 RNA 表現量較轉殖以 CaMV 35S 啟動子質體的
植株高。經 Bt 基因轉移之植株的生物檢定結果顯示,以 rbcS 啟動子為
轉殖質體的質株,殺小菜蛾效果較以 CaMV 35S 啟動子為轉殖質體的植株
高。目前轉殖三種基因的基因的三種轉殖植株均有再生。其中轉殖Bt及AF
基因之再生植物已完成生物檢定,轉殖MT基因已偵測有mRNA的存在,需再
做進一步生物檢定並探討轉殖植株後代之轉殖性狀的遺傳分析。

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.
URI: http://hdl.handle.net/11455/29259
Appears in Collections:園藝學系

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