Helicobacter pylori with separate β- and β′-subunits of RNA polymerase is viable and can colonize conventional mice

被引:15
作者
Raudonikiene, A
Zakharova, N
Su, WW
Jeong, JY
Bryden, L
Hoffman, PS
Berg, DE
Severinov, K
机构
[1] Rutgers State Univ, Waksman Inst, Piscataway, NJ 08854 USA
[2] Washington Univ, Sch Med, Dept Genet, St Louis, MO 63110 USA
[3] Dalhousie Univ, Dept Microbiol & Immunol, Halifax, NS B3H 4H7, Canada
[4] Rutgers State Univ, Dept Genet, Piscataway, NJ 08854 USA
[5] Washington Univ, Sch Med, Dept Mol Microbiol, St Louis, MO 63110 USA
关键词
D O I
10.1046/j.1365-2958.1999.01336.x
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
The genes encoding the beta- and beta '-subunits of RNA polymerase (rpoB end rpoC respectively) are fused as one continuous open reading frame in Helicobacter pylori and in other members of this genus, but are separate in other bacterial taxonomic groups, including the closely related genus Campylobacter. To test whether this beta-beta ' tethering is essential, we used polymerase chain reaction-based cloning to separate the rpoB and rpoC moieties of the H. pylori rpoB-rpoC fusion gene with a non-polar chloramphenicol resistance cassette containing a new translational start, and introduced this construct into H. pylori by electrotransformation. H. pylori containing these separated rpoB and rpoC genes in place of the native fusion gene produced non-tethered beta and beta ' RNAP subunits, grew well in culture and colonized and proliferated well in conventional C57BL/8 mice. Thus, the extraordinary beta-beta ' tethering is not essential for H. pylori viability and gastric colonization.
引用
收藏
页码:131 / 138
页数:8
相关论文
共 27 条
[1]   CONTROL OF TRANSCRIPTION TERMINATION [J].
ADHYA, S ;
GOTTESMAN, M .
ANNUAL REVIEW OF BIOCHEMISTRY, 1978, 47 :967-996
[2]   ADAPTIVE MUTATION AND COCOLONIZATION DURING HELICOBACTER-PYLORI INFECTION OF GNOTOBIOTIC PIGLETS [J].
AKOPYANTS, NS ;
EATON, KA ;
BERG, DE .
INFECTION AND IMMUNITY, 1995, 63 (01) :116-121
[3]   Analyses of the cag pathogenicity island of Helicobacter pylori [J].
Akopyants, NS ;
Clifton, SW ;
Kersulyte, D ;
Crabtree, JE ;
Youree, BE ;
Reece, CA ;
Bukanov, NO ;
Drazek, ES ;
Roe, BA ;
Berg, DE .
MOLECULAR MICROBIOLOGY, 1998, 28 (01) :37-53
[4]   EXTENSIVE HOMOLOGY AMONG THE LARGEST SUBUNITS OF EUKARYOTIC AND PROKARYOTIC RNA-POLYMERASES [J].
ALLISON, LA ;
MOYLE, M ;
SHALES, M ;
INGLES, CJ .
CELL, 1985, 42 (02) :599-610
[5]  
Ausubel FM., 1993, Current Protocols in Molecular Biology
[6]  
Berg DJ, 1998, AM J PATHOL, V152, P1377
[7]  
Burgess RR, 1976, RNA POLYMERASE, P69
[8]  
CESAREO SD, 1992, FEMS MICROBIOL LETT, V99, P15, DOI 10.1016/S0378-1097(01)00133-1
[9]   THE EVOLUTIONARY HISTORY OF THE 1ST 3 ENZYMES IN PYRIMIDINE BIOSYNTHESIS [J].
DAVIDSON, JN ;
CHEN, KC ;
JAMISON, RS ;
MUSMANNO, LA ;
KERN, CB .
BIOESSAYS, 1993, 15 (03) :157-164
[10]   THE EVOLUTION OF THE HISTIDINE BIOSYNTHETIC GENES IN PROKARYOTES - A COMMON ANCESTOR FOR THE HISA AND HISF GENES [J].
FANI, R ;
LIO, P ;
CHIARELLI, I ;
BAZZICALUPO, M .
JOURNAL OF MOLECULAR EVOLUTION, 1994, 38 (05) :489-495