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題 名 | 荔枝露疫菌粒線體核酸探針之選殖與分析=Cloning and Analysis of mtDNA Probes of Peronophythora Litchii |
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作 者 | 陳隆鐘; 賴學珍; 李崇正; 鍾依紋; 葉瑩; 安寶貞; | 書刊名 | 植物保護學會會刊 |
卷 期 | 40:2 1998.06[民87.06] |
頁 次 | 頁95-108 |
分類號 | 434.281 |
關鍵詞 | 荔枝露疫菌; 粒線體核酸; 核酸探針; 墨點漬染法; Peronophythora litchii; mtDNA; DNA probes; Dot blot hybridization; |
語 文 | 中文(Chinese) |
中文摘要 | 利用三種內限制酵素 EcoR I、Hinc II 及( EcoR I + Sac I )剪切荔枝露疫菌PL024菌株之粒線體核酸,將所得核酸片段與pUC 18載體黏合並進行選殖,結果選殖到4個選殖株A2-3、HC3、HC9及ES6,其插入片段大小各為0.6 kb、0.4 kb、0.65 kb及0.6 kb,分別製備為核酸探針。分別以墨點漬染法與25株荔枝露疫菌粒線體核酸進行雜合反應,均有雜合交互作用。另以17株Phytophthora菌株及Pythium splendens的全量核酸與核酸探針A2-3、HC3、HC9及ES6進行雜合反應,結果HC3、HC9、ES6核酸探針與Pythium splendens皆無雜合交互作用;ES6 核酸探針與17株Phytophthora菌株沒有任何雜合交互作用,A2-3核酸探針除Phytophthora cryptogea外都有雜合交互作用,顯示ES6核酸探針比A2-3、HC3及HC9核酸探針對 荔枝露疫菌較具有專一性。經墨點漬染測試,ES6 核酸探針的靈敏度可測到100 fg濃度的露疫菌粒線體核酸。 |
英文摘要 | The mtDNA of Peronophythora litchii PL024 was digested with EcoR I,Hinc Ⅱ and EcoR I/Sac I,and the DNA fragments were ligated with vector pUC18. And then the recombinated plasmids were transformated into E. coli DH5 α. There were four recombinat clones, designed as A2-3, HC3, HC9 and ES6 respectively. Their sizes are 0.6, 0.4, 0.65 and 0.6kb. Probes A2-3, HC3, HC9 and ES6 showed strong signals in dot blot hybridization with 25 P. litchii isolates. Four probes hybridized with 17 Phytophthora isolates and Pythium splendens, and none of any hybridization signals were detected except probe A2-3. None of any hybridization signals was detected as ES6 probe hybridized with 17 Phytophthora isolates. Probe A2-3 hybridized with Phytophthora isolates except Phytophthora cryptogea. Specificity of probe ES6 to P. litchii was higher than A2-3, HC3, HC9. In dot blotting analysis, a weak hybridization signals were shown when these four probes hybridized with 16 Pythium isolates, and ES6 probe was sensitive to mtDNA of Peronophythora litchii in concentration of 100 fg. |
本系統中英文摘要資訊取自各篇刊載內容。