Cloning and characterization of Rv0621 gene related to surfactant stress tolerance in Mycobacterium tuberculosis.
To understand how Mycobacterium tuberculosis (M. tuberculosis) could survive in human lung, Genomic expression library of M. tuberculosis in Escherichia coli (E. coli) had been prepared. Taking advantage of the genetic simplicity of E. coli and the functional conservation of some prokaryote proteins, a surfactant stress resistant gene Rv0621 was identified, which encodes a 37 kDa putative membrane protein. The E. coli colony with the partial Rv0621 gene insert, named S1, was able to grow in medium containing 0.4% sodium dodecyl sulfate, while the strain carried empty vector was unable to grow. The full length of the Rv0621 gene was then cloned into plasmid pET32a (+) expressed in E. coli BL21 (DE3). Using gas chromatographic-mass spectrometric (GC-MS), the fatty acid composition of the E. coli BL21 (DE3) carrying Rv0621-pET32a (+) and the E. coli BL21 (DE3) carrying empty vector pET32a (+) were compared. E. coli BL21 (DE3) carrying Rv0621-pET32a (+) contained more oleic acid. This suggests the gene may be involved in regulation of fatty acid synthesis and M. tuberculosis resistance to the surfactant defense of its host.
Dan
Liao
L
Institute of Modern Biopharmaceuticals, School of Life Sciences, Southwest University, Chongqing, People's Republic of China.
Jianping
Xie
X
Ruzhen
Guo
G
Honghai
Wang
W
D000222:Adaptation, Physiological / Q000187:drug effects*; D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000032:analysis / Q000737:chemistry / Q000235:genetics; D003001:Cloning, Molecular; D004269:DNA, Bacterial / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D005227:Fatty Acids / Q000032:analysis; D015964:Gene Expression Regulation, Bacterial / Q000187:drug effects; D015723:Gene Library; D005798:Genes, Bacterial; D006801:Humans; D050296:Microbial Viability / Q000187:drug effects; D008969:Molecular Sequence Data; D009169:Mycobacterium tuberculosis / Q000187:drug effects* / Q000235:genetics* / Q000254:growth & development; D010957:Plasmids / Q000235:genetics; D011994:Recombinant Proteins / Q000302:isolation & purification / Q000378:metabolism; D015183:Restriction Mapping; D017422:Sequence Analysis, DNA; D012967:Sodium Dodecyl Sulfate / Q000494:pharmacology; D013312:Stress, Physiological / Q000187:drug effects* / Q000235:genetics*; D013501:Surface-Active Agents / Q000494:pharmacology*; D014170:Transformation, Genetic / Q000187:drug effects
D001426:Bacterial Proteins; D004269:DNA, Bacterial; D005227:Fatty Acids; D011994:Recombinant Proteins; D013501:Surface-Active Agents; D012967:Sodium Dodecyl Sulfate