CAJ | 학술논문

利用自主分离的芽胞杆菌菌株TS01和15种芽胞杆菌(地衣芽胞杆菌(Bacillu.licheniformis)、枯草芽胞杆菌(B.subtilis)、短小芽胞杆菌(B.pumilus)、巨大芽胞杆菌B(.megaterium)、凝结芽胞杆菌(B.coagulans)、蜡状芽胞杆菌(B.cereus)、迟缓芽胞杆菌(B.lentus)、苏云金芽胞杆菌(B.thuringiensis)、嗜热脂肪芽胞杆菌(B.thermoaerophilus)、解淀粉芽胞杆菌(B.amyloliquefaciens)、环状芽胞杆菌(B.circulans)、球形芽胞杆菌(B.sphaericus)、侧胞短芽胞杆菌(Brevibacillus laterosporus)、多粘类芽胞杆菌(Paenibacillus polymyxa)和泛酸枝芽胞杆菌(Virgibacillus pantothenticus))模式菌种进行扩增核糖体DNA限制性分析(ARDRA).采用16S rDNA通用引物16S-27和16S-1525进行PCR扩增,16S rDNA扩增片段经6种限制性酶(Alu Ⅰ、TaqⅠ、Mse Ⅰ、Bst U Ⅰ、Hha Ⅰ和Tsp509 Ⅰ)酶切电泳,获得了TS01菌株的特征性ARDRA指纹图谱.ARDRA图谱通过GelcomparⅡ软件进行聚类分析(UPGMA),结果表明,菌株TS01和地衣芽胞杆菌处于同一分支,亲缘关系最近.ARDRA分析鉴定结果与实验室前期菌株TS01形态、生化鉴定和16SrDNA序列分析结果一致,TS01是1株地衣芽胞杆菌菌株,从而证明ARDRA技术在菌种水平上对芽胞杆菌TS01进行鉴别具有可靠性.
이용자주분리적아포간균균주TS01화15충아포간균(지의아포간균(Bacillu.licheniformis)、고초아포간균(B.subtilis)、단소아포간균(B.pumilus)、거대아포간균B(.megaterium)、응결아포간균(B.coagulans)、사상아포간균(B.cereus)、지완아포간균(B.lentus)、소운금아포간균(B.thuringiensis)、기열지방아포간균(B.thermoaerophilus)、해정분아포간균(B.amyloliquefaciens)、배상아포간균(B.circulans)、구형아포간균(B.sphaericus)、측포단아포간균(Brevibacillus laterosporus)、다점류아포간균(Paenibacillus polymyxa)화범산지아포간균(Virgibacillus pantothenticus))모식균충진행확증핵당체DNA한제성분석(ARDRA).채용16S rDNA통용인물16S-27화16S-1525진행PCR확증,16S rDNA확증편단경6충한제성매(Alu Ⅰ、TaqⅠ、Mse Ⅰ、Bst U Ⅰ、Hha Ⅰ화Tsp509 Ⅰ)매절전영,획득료TS01균주적특정성ARDRA지문도보.ARDRA도보통과GelcomparⅡ연건진행취류분석(UPGMA),결과표명,균주TS01화지의아포간균처우동일분지,친연관계최근.ARDRA분석감정결과여실험실전기균주TS01형태、생화감정화16SrDNA서렬분석결과일치,TS01시1주지의아포간균균주,종이증명ARDRA기술재균충수평상대아포간균TS01진행감별구유가고성.
Bacillus TS01 is a functional strain separated by authors'laboratory.TS01 strain and other 15 Bacillus type strains (B.licheniformis,B.subtilis,B.pumilus,B.megaterium,B.coagulans,B.cereus,B.lentus,B.thuringiensis,B.thermoaerophilus,B.amyloliquefaciens,B.circulans,B.sphaericus,Brevibacillus laterosporus,Paenibacillus polymyxa and Virgibacillus pantothenticus)were used to identify the differentiation by amplified ribosomal DNA restriction analysis(ARDRA).16S rDNA Wag amplified by the general primer pair 16S-27 and 16S-1525.The 16S rDNA PCR amplicon(about 1.5 kb)was digested by six restriction enzymes(Alu Ⅰ,Taq Ⅰ,Mse Ⅰ,Bst U Ⅰ,Hha Ⅰ and Tsp509 Ⅰ).And the characteristic ARDRA fingerprint of TS01 was obtained.The ARDRA patterns were analyzed by Gelcompar Ⅱ software using UPGMA method.And the homology tree based on ARDRA analysis showed that TS01 strain belonged to B.licheniformis,which was identical to the identification results of TS01 by using morphology,biochemistry and 16S rDNA sequence analysis.And the results showed that the established technique of ARDRA analysis has the dependability for the identification of TS01 strain on the specific level.