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標題: 菸草基因NbTRXh1影響竹嵌紋病毒移動的研究
The study of the gene NbTRXh1 from Nicotiana benthamiana involving in the movement of Bamboo mosaic virus
作者: 陳惠婷
Chen, Hui-Ting
關鍵字: BaMV;竹嵌紋病毒;movement;host factor
出版社: 生物科技學研究所
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竹嵌紋病毒(Bamboo mosaic virus, BaMV)為一個單股正極核糖核酸病毒,隸屬於Flexiviridae 科Potexvirus屬。病毒在複製、移動的過程中,往往會有許多的宿主基因參與並扮演不同角色。為了找出與竹嵌紋病毒生活週期有關的宿主基因,在先前研究中,本實驗室利用cDNA amplified fragment polymorphism (cDNA-AFLP ) 技術在菸草植物 ( Nicotiana benthamiana) 中篩選出了九十個在竹嵌紋病毒感染後具有差異性表現的基因片段。其中一個基因ACGT4在經過5'' RACE 延長5''端後分析其序列,我們發現這個基因ACGT4可能編碼出硫氧還原蛋白h (thioredoxin h protein)。根據先前的報導得知,硫氧還原蛋白h為一個可以還原雙硫鍵的小分子蛋白,它具有一個保留性的活化區WCGPC。有一些硫氧還原蛋白h亦被指出具有在細胞間移動的能力。為了瞭解ACGT4這個可能是硫氧還原蛋白h的基因會對竹嵌紋病毒產生什麼影響,我們利用病毒誘導基因靜默技術(VIGS)來抑制ACGT4在菸草中的表現,結果發現在ACGT4表現被抑制的植物中,竹嵌紋病毒的鞘蛋白在接種後的第五天有較高的累積量,但這樣的結果並未出現於ACGT4表現被抑制的植物原生質體中。另外我們也發現當菸草中的ACGT4表現量被抑制時,菸草植物較容易被竹嵌紋病毒感染,且竹嵌紋病毒會具有較高的移動效率。因此ACGT4可能會影響竹嵌紋病毒在植物中的移動但不影響其複製。至於ACGT4影響竹嵌紋病毒的確切機制是什麼,則有待進一步的研究確認。

Bamboo mosaic virus (BaMV) is a single-stranded positive-sense RNA virus. It belongs to the Potexvirus genus of Flexiviridae. In general, RNA virus encodes a limit number of genes and requires a few different host factors to fulfill its infection cycle such as translation, replication, and movement. Previously, our laboratory had used the cDNA-amplified fragment polymorphism (cDNA-AFLP) technique identifying about 90 differentially expressed gene fragments from Nicotiana benthamiana plants upon BaMV infection. One of the genes ACGT4 may encode a thioredoxin h protein after sequence analysis of the full-length cDNA derived from the clones of the 5''-rapid amplification of complementary DNA ends (5'' RACE). The thioredoxin h proteins are a group of small proteins with a conserved WCGPC active site conferring a disulfide reductase activity. Some of the thioredoxin h proteins have been reported to have the ability to move among cells. To inspect how the ACGT4 involves in the infection cycle of BaMV in N. benthamiana plant. I use the virus induced gene silencing (VIGS) system to knock down the expression level of ACGT4 in N. benthamiana plant and then inoculate with BaMV. Results show that the accumulation of BaMV coat protein is increased in the knockdown plant at 5 dpi compared to that of control plant. However, I do not find any significant difference between the knockdown and control protoplasts at 24 hpi. I also find that BaMV is more efficient in infection and movement in ACGT4-knockdown plant than those in control plants. Overall of these results suggest that the product of ACGT4 gene may play a role in restricting BaMV movement rather in replication. The mechanistic involvement of ACGT4 in BaMV movement is still required for further analysis.
其他識別: U0005-1208201120452900
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