枯草桿菌F29-3全基因體定序與表現型分析

Whole-Genome Sequencing and Phenotypic Analysis of Bacillus subtilis F29-3

張凱欣

枯草桿菌F29-3 ； 全基因體定序 ； 生長特性 ； Bacillus subtilis F29-3 ； whole genome sequencing ； growth characteristics ； phenotype MicroArray

長榮大學生物科技學系（所）學位論文

2016年

碩士

許鴻猷

繁體中文

枯草桿菌為生長於土壤表層的革蘭氏陽性桿菌，具有產生內胞子的能力，對人類不具致病性。枯草桿菌屬的細菌，由於具有產生抗生素的能力，因此常被應用於食品、醫藥、與植物病害的生物防治。枯草桿菌F29-3是絲核菌類真菌Rizoctonia solani拮抗菌株，篩選自台中市豐原區。若將此細菌培養於Difco Sporulation Medium培養基中，經隔夜培養其可產生內胞子，培養於nHA固態培養基上菌落邊緣平整。多種桿菌屬細菌會產生桿菌素(Bacilysin)，它是由L-alanine與L-anticapsin這兩種胺基酸組成的抗生素，可以抑制酵母菌與金黃色葡萄球菌的生長，除桿菌素外枯草桿菌F29-3可產生豐原素(Fengycin)，豐原素是由枯草桿菌F29-3所生產的脂胜肽抗生素，其結構包含由十個胺基酸所組成的環狀胜肽鏈及一長碳鏈之脂肪酸。豐原素的胜肽鏈是經由非核醣體胜肽合成酶 (Nonribosomal peptide synthetases; NRPS)合成，在自然界中，非核醣體胜肽合成酶系統參與多種抗生素的合成。為了更加了解枯草桿菌F29-3，我們以霰彈槍定序法進行枯草桿菌F29-3的全基因體定序，由定序結果我們發現枯草桿菌F29-3基因體長度為4,195,514 bp，其G+C%為44.53%，共包含4,536個ORFs、10個16S/23S/5S rRNA operons、87個 tRNA genes、3個 NRPS loci (surfactin, fengycin, bacilysin)、2個 PKS (polyketide synthase;聚酮合成酶) loci與10個prophage。除了基因體定序外，我們也將枯草桿菌F29-3培養於960種不同營養成份以測定其使用這些養份的生長情形。這些結果將有助於我們更加了解枯草桿菌F29-3，並可應用於不同領域。

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