Identification of unique genetic markers in Rv0927c among Mycobacterium tuberculosis W-Beijing strains.
Fifty-six clinical isolates of Mycobacterium tuberculosis were analyzed by spoligotyping to determine the prevalence of W-Beijing strains. Forty-nine of the 56 isolates belonged to W-Beijing strains and 7 isolates were non-Beijing strains. Comparative two-dimensional gel electrophoresis analysis of protein patterns between the W-Beijing and non-Beijing strains identified a unique protein Rv0927c that is absent in the former but present in the latter and the reference strain M. tuberculosis H37Rv. Compared with 7 non-Beijing clinical isolates and H37Rv, all 49 W-Beijing strains had two characteristic mutations, a deletion of AGC at nucleotide position 421 of Rv0927c gene encoding a putative short dehydrogenase/reductase, causing deletion of serine codon at amino acid position 141 and a -127 G-->A mutation in Rv0927c-pstS3 intergenic region, resulting in failure to express Rv0927c. Western blot analysis indicated that polyclonal antibody raised against H37Rv Rv0927c overexpressed in Escherichia coli reacted with non-Beijing strains and H37Rv but not W-Beijing strains. Characteristic mutations of Rv0927c that are present in W-Beijing strains can be used as a novel genetic marker for rapid molecular typing of M. tuberculosis W-Beijing strains.
Jiang
Xin
X
State Key Laboratory of Genetic Engineering, Institute of Genetics, School of Life Science, Fudan University, Shanghai 200433, PR China.
Zhang
Ying
Y
Zhang
Wenhong
W
Gao
Feng
F
Jin
Jialin
J
Zhang
Xuelian
X
Lv
Chong
C
Gao
Qian
Q
Chen
Jing
J
Wang
Honghai
H
D000907:Antibodies, Bacterial / Q000276:immunology; D001426:Bacterial Proteins / Q000032:analysis / Q000235:genetics*; D015153:Blotting, Western; D004269:DNA, Bacterial / Q000235:genetics; D021901:DNA, Intergenic; D015180:Electrophoresis, Gel, Two-Dimensional; D015870:Gene Expression; D005819:Genetic Markers; D006801:Humans; D007106:Immune Sera / Q000276:immunology; D009169:Mycobacterium tuberculosis / Q000737:chemistry / Q000145:classification* / Q000235:genetics*; D010088:Oxidoreductases / Q000235:genetics; D017354:Point Mutation; D016133:Polymerase Chain Reaction; D020543:Proteome / Q000032:analysis; D017422:Sequence Analysis, DNA; D017384:Sequence Deletion; D014376:Tuberculosis / Q000382:microbiology
D000907:Antibodies, Bacterial; D001426:Bacterial Proteins; D004269:DNA, Bacterial; D021901:DNA, Intergenic; D005819:Genetic Markers; D007106:Immune Sera; D020543:Proteome; D010088:Oxidoreductases