Please use this identifier to cite or link to this item: http://hdl.handle.net/11455/21424
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dc.contributor.advisorWen-Hwei Hsuen_US
dc.contributor.advisor許文輝zh_TW
dc.contributor.author陳昶霖zh_TW
dc.contributor.authorChen, Chang-Linen_US
dc.date2004zh_TW
dc.date.accessioned2014-06-06T07:15:47Z-
dc.date.available2014-06-06T07:15:47Z-
dc.identifier.urihttp://hdl.handle.net/11455/21424-
dc.description.abstractP1201 is a lytic corynephage of Corynebacterium glutamicum NCHU 87078. The linear double—stranded DNA genome was 70,579 bp, with 3'—protruding cohesive ends of ten nucleotides and an overall G+C content of 50.9%. Analysis of complete genome with cumulative GC skew diagrams, the low point, near the left end, with short repeats and lacking ORFs, might be the site of replication origin. One hundred and fifteen open reading frames were identified. The genomic organization of the putative coding regions was very similar to that of mycobacteriophage. Four tRNA—like sequences (two tRNA—Arg, one tRNA—Trp, and one tRNA—Gly) were found, in which two tRNA—Arg genes for codons AGA and AGG are rare in C. glutamicum. The genes required for rgulation, packaging, head assembly, tail assembly, lysis, replication in P1201 genome were homologous to mycobacteriophage Omega, mycobacteriophage Che8, actinophage VWB, mycobacteriophage CJW1, mycobacteriophage Bxz2, mycobacteriophage Barnyard and bacteriophage SPβ respectively. Corynephage P1201 had three inteins which integrate into ORF 27 (thymidylate synthase), ORF 34 (terminase), and ORF 87 (RNR α subunit), respectively. To our knowledge, this is the first report that intein has been integrated into thymidylate synthase gene. The existence of three inteins in corynephage genome may play important roles in the horizontal exchange of genetic material during evolution. The mosaic—like genome of P1201 indicated the occurrence of horizontal gene exchange among corynephages, mycobacteriophages, Corynebacteriaceae, and Mycobacteriaceae in corynephage P1201 evolution. We suggested that corynephages and mycobacteriophages come from the same pool.en_US
dc.description.abstractCorynephage P1201 是從味精生產菌 Corynebacterium glutamicum NCHU 87078 分離出來的溶裂性噬菌體。基因組由 70,579 bp 雙股 DNA 所組成,染色體 3’ 末端凸出是由 10 個核苷酸所組成。整個基因組的 G+C 含量為 50.9%。利用 GC skew 分析其差值最低點位於左邊端點,此區域可能是複製的啟始區;最高點在 39 kb 左右的位置,可能是複製的終止區。利用生物資訊分析其基因組後,推測噬菌體 P1201 含有 115 個基因,包括 111 個開放讀碼區和 4 個 tRNA 基因 (兩個 tRNA—Arg,一個 tRNA—Trp 和一個 tRNA—Gly)。這兩個 tRNA—Arg 所對應的密碼為 AGA 和 AGG,目前已知帶有這兩組反密碼的 tRNA 在 C. glutamicum 中是較稀少的。噬菌體 P1201 基因組之功能由 5’ 端到 3’ 依序為:Regulation—Packaging—Head assembly—Tail assembly—Lysis—Replication—Regulation,這些區域的蛋白質分別和 mycobacteriophage Omega、mycobacteriophage Che8、actinophage VWB、mycobacteriophage Cjw1、mycobacteriophage Bxz2、mycobacteriophage Barnyard 及 bacteriophage SPβ 有很高的相似度。噬菌體 P1201 含有三個 intein,分別插在 ORF 27 (thymidylate synthase)、ORF 34 (terminase) 及 ORF 87 (RNR α subunit) 之中。據我們所知,這是第一個發現有 intein 插入 thymidylate synthase 基因的例子。在基因組演化過程中,此三個 intein 可能會導致基因的水平轉移,而造成基因重組的變異。噬菌體 P1201 基因組呈現出高度馬賽克的結構,顯示此噬菌體在演化過程中,和 corynephages 與 mycobacteriophages 噬菌體及 Corynebacteriaceae 與 Mycobacteriaceae 菌體之間存在基因水平轉移的現象,另一方面藉由分析噬菌體 P1201 的基因組可推測 corynephages 和 mycobacteriophages 是來自相同的生物庫。zh_TW
dc.description.tableofcontents目 錄 I 圖目錄 IV 表目錄 V 摘 要 VI Abstract VII 縮寫對照表 VIII 前 言 1 1. Corynebacterium glutamicum 1 1.1. C. glutamicum 及其親緣相近之菌種 1 1.2. C. glutamicum 在工業及學術上之應用 1 2. 噬菌體 3 2.1. 噬菌體的分類 3 2.2. 噬菌體在自然界所扮演的角色 4 2.3. 噬菌體的生活史 4 2.4. 噬菌體之基因體結構 6 2.5. 噬菌體的演化及基因轉移 7 2.6. 噬菌體在生技產業上的應用 9 2.7. Corynephage 及其相關噬菌體基因體的解碼 10 材料與方法 12 1. 藥品 12 2. 菌種、噬菌體、質體及培養基 12 3. 質體抽取 16 4. 噬菌體 DNA 的抽取 16 5. DNA 回收、接合及聚合酶鏈鎖反應 17 6. 蛋白質電泳分析 17 7. 轉形作用 17 8. 噬菌體 P1201 基因體之 DNA 序列分析 18 9. 噬菌體 P1201 基因體結構的分析 22 10. 噬菌體生長曲線的分析 23 結 果 24 1. P1201 的型態 24 2. 噬菌體 P1201 的生長曲線 24 3. P1201 基因組的分析 26 3.1. 基因的排列及其 ORF 的比對 29 3.2. tRNA 的基因序列 53 3.3. 構造基因的分析 56 3.4. GC skew 分析 60 討 論 62 1. Modular theory 在 P1201 的驗證 62 2. TMP 的修飾 66 3. 特殊 tRNA 的使用率 67 4. 噬菌體 P1201 中的 intein 基因 72 總 結 76 參考文獻 77zh_TW
dc.language.isoen_USzh_TW
dc.publisher分子生物學研究所zh_TW
dc.subjectphageen_US
dc.subject噬菌體zh_TW
dc.subjectCorynebacterium glutamicumen_US
dc.subject味精生產菌zh_TW
dc.titleGenomic Sequence and Analysis of Corynephage P1201en_US
dc.title噬菌體 P1201 基因組之解碼及分析zh_TW
dc.typeThesis and Dissertationzh_TW
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