Please use this identifier to cite or link to this item: http://hdl.handle.net/11455/13811
標題: 家禽里奧病毒基因體泳動圖譜之比較與 S1133 病毒株 S3 片段核酸序列之定性分析
Comparison of the migration pattern of avian reovirus genome and characterization of the nucleotide sequence of virus strain S1133 S3 fragment
作者: 吳婉瑜
Wu, Wan Yu
關鍵字: avian reovirus;家禽里奧病毒;migration pattern;dsRNA;cloning;sequencing cDNA;移動圖譜;雙股 RNA;選殖;定序;互補 DNA
出版社: 獸醫學系
摘要: 
家禽里奧病毒基因體包含十段雙股 RNA,此些片段主要分為三群,L、 M
、S。以 PAGE 分析十三株家禽里奧病毒,其中 S1133、2408、1733為疫
苗株,其餘為十株為野外分離株。結果顯示 S 群比 L 群有較高的變異性
。病毒基因體泳動圖譜與病毒的血清型或引發之病變間沒有相關性。純化
家禽里奧病毒 S1133 病毒株之雙股 RNA 以反轉錄鋂合成 cDNA。
terminal transferase 於 cDNA 加上 oligo(dC) 以 pstI site與
oligo(dG)-tailed pBR322 行連接作用,篩選得到十二個重組質體,次選
殖於 pUC18 質體。取 W1,含1 kb 之崁入 cDNA 經定序顯示此片段含有
一開放讀碼 852 個核甘酸,轉譯蛋白約 35.983 kDa。以點墨吸自漬証實
此 cDNA 片段源於家禽里奧病毒 S3 片段。此重組質體 W1 片段序列有
17 個 ATG,而此片段為哺乳動物里奧病毒基因體之特徵。而哺乳類里奧
病毒基因體 S4 啟始密碼位於 5''端第 32個位置而 W1 片段啟始密碼位
於 5''端第 24 個核蜬酸位置,推測重組質體可能是家禽里奧病毒基因體
S3 片段之全長。本實驗結果除進一步了解家禽里奧病毒基因體外,寄望
能運用於診斷上。

RNA isolate from ARV strain S1133 was adenylated wite poly(A)
polymerase.The poly(A)‧RNA was isolated by oligo(dT)-cellulose
chromatography and was reversed transcribed with M-MLV reverse
transcriptase. The resulting DNA was tailed with oligi(dC)
with terminal transferase and was cloned into oligo(dG)-tailed
pBR322 at the PstI site. W1 which contained 1-kb cDNA insert
was sequenced.Result show that this fragment contained an open
reading frame of 852 nucleotides,encoding a protein of 35.983
kDa.Dot-blot hybrydization results showed that the cDNA
fragment was located on the S3 segment of ARV.The amino acid
seqence of the putative protein was highly similar to the
seqence of the outer capsid protein,σ3,encoded by the S4
segment of mamalian reovirus (MRV) .Therefore,the ARV open
reading frame may also encode a protein of similar function.
The results obtained from this study wil be useful for the
understanding of ARV and for the diagnosis of the disease
caused by the pathogen.
URI: http://hdl.handle.net/11455/13811
Appears in Collections:獸醫學系所

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