A single band does not always represent single bacterial strains in denaturing gradient gel electrophoresis analysis

被引：202

作者：

Sekiguchi, H ^{[1
]}

Tomioka, N ^{[1
]}

Nakahara, T ^{[1
]}

Uchiyama, H ^{[1
]}

机构：

[1] Univ Tsukuba, Inst Appl Biochem, Tsukuba, Ibaraki 3058577, Japan

来源：

BIOTECHNOLOGY LETTERS | 2001年 / 23卷 / 15期

关键词：

bias; DGGE; microbial community; PCR; 16S rDNA;

D O I：

10.1023/A:1010517117046

中图分类号：

Q81 [生物工程学（生物技术）]; Q93 [微生物学];

学科分类号：

071005 ; 0836 ; 090102 ; 100705 ;

摘要：

DNA in a denaturing gradient gel electrophoresis (DGGE) band that could not be sequenced after recovery from the gel was cloned into a TA cloning vector and a library was constructed and then 13 clones randomly picked up from the library was sequenced. Although the excised DNA from the DGGE gel showed a single band, the library consisted of several different sequences phylogenetically. This phenomenon was also observed in several other DGGE bands. Therefore, this suggests that a single DGGE band does not always represent a single bacterial strain and a new bias for quantitative analyses based on band intensities has been identified.

引用

页码：1205 / 1208

页数：4

共 13 条

[1] PHYLOGENETIC IDENTIFICATION AND IN-SITU DETECTION OF INDIVIDUAL MICROBIAL-CELLS WITHOUT CULTIVATION [J].

AMANN, RI ;

LUDWIG, W ;

SCHLEIFER, KH .

MICROBIOLOGICAL REVIEWS, 1995, 59 (01) :143-169

[2] Denaturing gradient gel electrophoresis profiles of 16S rRNA-defined populations inhabiting a hot spring microbial mat community [J].

Ferris, MJ ;

Muyzer, G ;

Ward, DM .

APPLIED AND ENVIRONMENTAL MICROBIOLOGY, 1996, 62 (02) :340-346

[3] Microbial evolution, diversity, and ecology: A decade of ribosomal RNA analysis of uncultivated microorganisms [J].

Head, IM ;

Saunders, JR ;

Pickup, RW .

MICROBIAL ECOLOGY, 1998, 35 (01) :1-21

[4] BACTERIAL POPULATIONS IN SEA WATER AS DETERMINED BY DIFFERENT METHODS OF ENUMERATION [J].

JANNASCH, HW ;

JONES, GE .

LIMNOLOGY AND OCEANOGRAPHY, 1959, 4 (02) :128-139

[5] Multiple sequence alignment with Clustal x [J].

Jeanmougin, F ;

Thompson, JD ;

Gouy, M ;

Higgins, DG ;

Gibson, TJ .

TRENDS IN BIOCHEMICAL SCIENCES, 1998, 23 (10) :403-405

[6]

LIONEL R, 2000, RES MICROBIOL, V151, P167

[7] PROFILING OF COMPLEX MICROBIAL-POPULATIONS BY DENATURING GRADIENT GEL-ELECTROPHORESIS ANALYSIS OF POLYMERASE CHAIN REACTION-AMPLIFIED GENES-CODING FOR 16S RIBOSOMAL-RNA [J].

MUYZER, G ;

DEWAAL, EC ;

UITTERLINDEN, AG .

APPLIED AND ENVIRONMENTAL MICROBIOLOGY, 1993, 59 (03) :695-700

[8]

Muyzer G, 1998, ADV MOL ECOLOGY, P157

[9]

MYERS RM, 1985, NUCLEIC ACIDS RES, V13, P3131, DOI 10.1093/nar/13.9.3131

[10] THE NEIGHBOR-JOINING METHOD - A NEW METHOD FOR RECONSTRUCTING PHYLOGENETIC TREES [J].

SAITOU, N ;

NEI, M .

MOLECULAR BIOLOGY AND EVOLUTION, 1987, 4 (04) :406-425

← 1 2 →