| 蜡样芽孢杆菌ERIC-PCR分子分型方法的建立 |
| |
| 引用本文： | 王君,吴清平,吴克刚,陈谋通,张菊梅,郭伟鹏.蜡样芽孢杆菌ERIC-PCR分子分型方法的建立[J].现代食品科技,2013,29(7):1696-1701. |
| |
| 作者姓名： | 王君 吴清平 吴克刚 陈谋通 张菊梅 郭伟鹏 |
| |
| 作者单位： | 广东工业大学轻工化工学院;广东省微生物研究所 广东省华南应用微生物重点实验室-省部共建国家重点实验室 广东省菌种保藏与应用重点实验室 广东省微生物应用新技术公共实验室 |
| |
| 基金项目： | 广东省教育部产学研结合项目(2012B090400017);科技部国际科技合作专项(2013DFH30070) |
| |
| 摘 要： | 为建立简单快速的蜡状芽孢杆菌(Bacillus cereus)基因间重复共有序列PCR(ERIC-PCR)的分子分型方法,该研究以B.cereusCMCC 63303的基因组DNA为模板,并采用正交设计L16(45)对影响ERIC-PCR反应体系的五个因素(模板DNA、Mg2+、dNTP、TaqDNA聚合酶、引物)在四个浓度水平上进行优化,并通过单因素试验确定最佳退火温度,最后以优化后的反应体系对50株B.cereus分离株进行ERIC-PCR分子分型和聚类分析验证其稳定性和分型效果。结果显示应用建立的ERIC-PCR反应体系对50株分离株扩增均得到了9-17条、大小在200-4000 bp之间的条带,并可将其分为47个型,且分辨力达到0.996,具有较高稳定性和分辨能力。表明ERIC-PCR技术对B.cereus分型,具有简便和分辨力高等优点,可用于食源性B.cereus分离株的多样性研究。
|
| 关 键 词： | 蜡样芽孢杆菌 ERIC-PCR 分子分型 遗传多样性分析 |
| 收稿时间： | 4/7/2013 12:00:00 AM |
Establishment of ERIC-PCR Molecular Typing Method for Bacillus cereus |
| |
| WANG Jun,WU Qing-ping,WU Ke-gang,CHEN Mou-tong,ZHANG Ju-mei and GUO Wei-peng.Establishment of ERIC-PCR Molecular Typing Method for Bacillus cereus[J].Modern Food Science & Technology,2013,29(7):1696-1701. |
| |
| Authors: | WANG Jun WU Qing-ping WU Ke-gang CHEN Mou-tong ZHANG Ju-mei and GUO Wei-peng |
| |
| Affiliation: | 1.Faculty of Chemical Engineering and Light Industry,Guangdong University of Technology,Guangzhou 510006,China)(2.Guangdong Institute of Microbiology,South China State Key Laboratory of Applied Microbiology(Ministry-Guangdong Province Jointly Breeding Base),Guangdong Provincial Key Laboratory of Microbial Culture Collection and Application;Guangdong Open Laboratory of Applied Microbiology,Guangzhou 510070,China) |
| |
| Abstract: | An efficient enterobacterial repetitive intergenic consensus sequence (ERIC-PCR) molecular typing method was established for Bacillus cereus. Based on the template of B. cereus CMCC 63303 genomic DNA, the ERIC-PCR amplification system in four levels of five factors (DNA template, Mg2+, dNTP, Taq DNA polymerase and primer) was optimized by orthogonal design L16(45) .Then the effects of annealing temperature on the amplification system was discussed through single factor experiment and the optimal reaction condition were determined. Fifty B. cereus strains were typed using the optimal reaction system and cluster analysis. The ERIC-PCR results exhibited better discriminative results in molecular typing with discrimination index of 0.996. B. cereus strains were grouped into 47 types. The ERIC-PCR system was an efficient method for typing and tracking analyses. It was suitable for genetic diversity analysis of foodborne B. cereus strains. |
| |
| Keywords: | bacillus cereus ERIC-PCR molecular typing genetic diversity analysis |
| 本文献已被 CNKI 等数据库收录！ |
| 点击此处可从《现代食品科技》浏览原始摘要信息 |
|
点击此处可从《现代食品科技》下载全文 |
|
