题目：北极产碱性蛋白酶菌株VV52的分类鉴定.docVIP

题目：北极产碱性蛋白酶菌株VV52的分类鉴定 院（系） 化工学院 专 业 生物技术 届 别 2012届 学 号 0826321023 姓 名 林源清 指导老师 陈明霞 华侨大学教务处印制 2012年 05 月 北极产碱性蛋白酶菌株VV52的分类鉴定 摘要 菌株VV52是从北极水样中分离得到的产碱性蛋白酶细菌，本论文采用16S rNA基因 PCR-RFLP（限制性酶切多态性）方法对菌进行分型，进行16S rNA基因序列测定、Gene bank数据库blast分析通过DNAMAN软件系统进化Stenotrophomonas rhizophila(嗜根寡养单胞菌i-C15:0 (28.43%)和a-C15：0 （16.12%）占有绝对优势；运用高效液相色谱法（HPLC）对细菌DNA中G+C mol%的测定结果表明该菌株具有高的G+C mol%含量，其含量为64.2%；形态及生理生化性状研究显示菌株为直杆状，革兰氏阴性，The strain VV52 isolated from the Arctic waters was one of the alkaline protease producing bacteria. Firstly, it was identificated usine 16S rRNA gene PCR-RFLP (restriction enzyme polymorphism) and 16S rRNA gene sequence analysis methods. The result showed that strain VV52 was Stenotrophomonas rhizophila-like species. To further identify this strain, cellular fatty acid was detected by gas chromatorgaphy (GC) methods. The result showed that there were 17 kinds of main cellular fatty acid, iso-branched i-C15:0 and anteiso- branched i-C15:0 were the predominant fatty acid component and constituted 28.43% and 16.12% respectively. Then, we detected the G+C content of the DNA by high performance liquid chromatography (HPLC). The result showed that strain VV52 had high G+C content (64.2%). Study on the morphological, biochemical and physiological characteristics showed that?strain?VV52 is straight?rod-shaped,?Gram-negative,?no capsule; oxidase, urease and protease positive, DNase negative; hydrolyze?aescin?glycosides but not acetamide,?reduce nitrate,?did not produce? hydrogen sulfide; utilize mannitol,?xylose, trehalose, glucose,?maltose?and?sucrose, but not?lactose, galactose, fructose,?mannose, adonitol?and rhamnose. Keywords: 16S rRNA gene, RFLP, G+C content, cellular fatty acid analysis, bacterial classification and identification 目录