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標題: 立枯絲核菌線形質體pCHU352之定序及分析
Sequencing and analysing of linear plasmid pCHU352 of Rhizoctonia solani
作者: 隋孟君
Sui, Meng-Jiun
關鍵字: Rhizoctonia solani
linear plasmid
出版社: 植物學系
摘要: 本研究是針對立枯絲核菌第四融合群分離株CHU352的線形質體pCHU352進 行分析及定序。立枯絲核菌CHU352有三種同為2.6kb大小的線形質體,分 別命名為pCHU352-1、pCHU352-2和pCHU352-3,其中pCHU352-2中央區域 的1364bps核酸序列已被定序出來。經過DNASIS軟體分析後可得到 13組 inverted repeat sequences以及一段大小為418bps的open reading frame,此段核酸序列與Hongo等人於1994發表之研究結果具有高達95.7% 的同源性(Homology)。此外,將1364bps核酸與周氏之論文結果比對,亦 有60.2%的同源性。根據鹼性電泳的結果可證實pCHU352的末端應為髮夾狀 結構(hairpin loop structure),與Miyashita等人的研究結果相同。然 而,在膠體電泳分析中,二倍體的存在方面,也經由實驗證明二倍體並不 是在線形質體複製時所造成的,並非如Miyashita等人所提出之對於二倍 體存在的解釋。
Prvious studies had indicated that Rhizoctonia solani isolate CHU352 of anastomosis group 4, possesses three linear mitochon- drial DNA plasmids (pCHU352-1, -2 and -3). These linear DNA plasmids have similar size (2.6 kb). 1364 bps nucleotide sequence in the central region of pCHU352-2 has been determined. The sequencing data showed that there were 13 inverted repeat sequences over the entire sequenced region and 1 ORF (open reading frame) which contained 418 bps. The ORF homology between pCHU352 and pRS64 (Hongo et al., 1994) is 95.7%. The nucleotide sequence homology between pCHU352HH and pCHU341 (Chow, 1994) is 60.2%. The length of both terminal fragments that were generated by restriction endonuclease digestion was doubled under the denaturation condition, indicating that the linear plasmid DNAs have hairpin loops at the both termini. The formation of dimeric form is not according to the model of replication proposed by Miyashita et al..
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