末端限制性片段长度多态性技术_T_省略_微生物群体上的应用与技术优化_李献梅.pdfVIP

2009 , 14( 4) : 1- 9 J ournal o f China A g ricultur al U niv ersity ( T-RFLP) 李献梅 王小芬 崔宗均* ( , 100193) 末端限制性片段长度多态性技术( T- RFL P) 是以荧光标记引物P CR 为基础, 根据末端限制性片段长度区 分出微生物 体组成的一种微生物 体图谱法本文综述了该方法在 体微生物分析上的应用及DN A 提取 PCR 扩增酶切和数据分析等步骤的技巧与优化, 总结了科学应用该技术的要求以及和多重 PCRgyrB 基因等相 结合的观点 T- RF LP ; 微生物 体; 图谱分析; PCR; 克隆 Q 936 1007- 4333( 2009) 04-000 1- 09 A Applications and optimizations of T-RFLP infingerprinting environmental microbial populations * LIXian-mei, WAN Xiao-fen, CUIZong-jun ( College of Agronomy and Biot echnology Cent er of Biomas s Engineering , China Agricult ural Univers it y , Beijing , 100 193, China) Abstract Terminal re stric tio n fra gment leng th polymorphis m ( T- RFLP) is one o f micro bia l popula tio n fing erprinting me tho ds tha t is ba sed o n labele d primers- PCR to ide ntify microbial co mposition a ccording to diffe re nt terminal re stric tio n frag me nt leng ths . The applic ations and te chnica l o ptimizations w e re o ve rv iew ed o f T- RFLP thro ugh the pro cedures of DNA preparation, PCR, enzyme dig estion and da ta analysis . The sc ientific applic ation re quire me nts and new view s of combination w ith multiple- PCR o r g yr B g e ne w e re dra w n. Key words T- RFLP; microbial po pulation; fing erprinting ana lysis; PCR; clone libra ry RFLP , [ 1] , ) ) ) D GGE T GGESSCPFISH ) ) ) , [ 2- 6] 1 T- RFLP ( t er minal- T- RF L P r