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題 名 | 利用PCR-RFLP技術鑑定五種核多角體病毒臺灣分離株=Detection and Identification of Five Nucleopolyhedrovirus Isolates from Taiwan by PCR-RFLP |
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作 者 | 高穗生; 靳子蓉; 王重雄; | 書刊名 | 中華昆蟲 |
卷 期 | 20:1 2000.03[民89.03] |
頁 次 | 頁23-35 |
分類號 | 387.7 |
關鍵詞 | 核多角體病毒; 多角體蛋白基因; 聚合酶鏈鎖反應; 限制酶片段多型性; Nucleopolyhedrovirus; NPV; Polymerase chain reaction; PCR; Polyhedrin gene; Polh; Restriction fragment length polymorphisms; RFLP; |
語 文 | 中文(Chinese) |
中文摘要 | 對甜菜夜蛾(Spodoptera exigua)、斜紋夜蛾(S. litura)、榕樹透翅毒蛾(Perina nuda)、木毒蛾(Lymantria xylina)核多角體病毒及苜蓿夜蛾(Autographa californica)核多角體病毒台灣分離株等五種本土性核多角體病毒進行鑑定與分析的研究。經EcoRI 限制[西每]圖譜分析,證實此五種核多角體病毒的基因組DNA 各具有不同的EcoRI 限制[西每]圖譜,因此以EcoRI 限制[西每]圖譜分析法即可達到區分此五種本土性核多角體病毒的效果。但為求更進一步改善及提高對此五種本土性核多角體病毒偵測的靈敏度,故選擇以聚合[西每]鏈鎖反應(PCR)增幅多角體蛋白基因片段(約680bp),其增幅的片段除定序外,再與BsuRI 、BspHI 、BsiWI 、HpaII 、MseI 和TaqI 等限制[西每]進行限制[西每]片段多型性(RFLP)分析。其結果顯示此五種核多角體病毒經BsuRI 限制[西每]切割後即呈現不同的剖面,所以各核多角體病(NPV)之間的差異性可明顯地分辨開來。另 BspHI 、BsiWI 、HpaII 、MseI 和TaqI 之限制[西每]圖譜亦可輔助鑑定此五種核多角體病毒。因此PCR-RFLP 分析技術將可提供一套快速且靈敏的偵測系統以鑑定此五種本土性核多角體病毒。 |
英文摘要 | Five nucleopolyhedrovirus (NPV) isolates from Taiwan: Spodoptera exigua nucleopolyhedrovirus (SpeiNPV), Spodoptera litura NPV (SpltNPV), Perina nuda NPV (PenuNPV), Lymantria xylina NPV (LyxyNPV) and a isolate of Autographa californica NPV (AcMNPV-TWN4) from S. exigua were used for this study. EcoR Ⅰprofiles of genomic DNAs of five NPVs gave a clear pattern for identification, this result showed that EcoR Ⅰprofiles were sufficient to identify these NPVs. A polymerase chain reaction-restriction fragment length polymorphisms (PCR-RFLP) technique was used for rapid detecting and identifying NPVs which were even at low concentration. A set of primers, designated as primers 35 and 36 (Chou et al., 1996) was used for amplification of polyhedrin gene from five NPVs by PCR and the size of amplicons was 680 bps. These polyhedrin gene fragments were sequenced and were then digested with BsuR Ⅰ, BspH Ⅰ, BsiW Ⅰ,Hpa Ⅱ, Mse Ⅰand Taq ⅠDNA restriction endonuclease, respectively. The BsuR Ⅰ patterns of amplicons gave a completely different pattern from each other, and the profiles of other selected restriction enzymes could be regarded as a complement data for Bsu RI for detection and identification. It is suggested that PCR-RFLP technique is a feasible method for rapid detection and identification of five NPVs isolated from Taiwan. |
本系統中英文摘要資訊取自各篇刊載內容。