Cloning and expression of the Mycobacterium bovis BCG gene for extracellular alpha antigen.
The gene for the extracellular alpha antigen of Mycobacterium bovis BCG was cloned by using a single probe restricted to G or C in the third position. This technique should have great potential for the isolation of mycobacterial antigen genes. The gene analysis revealed that the alpha antigen gene encoded 323 amino acid residues, including 40 amino acids for signal peptide followed by 283 amino acids for mature protein. This is the first report on the structure of the mycobacterial signal peptide. The promoter-like sequence and ribosome-binding site were observed upstream of the open reading frame. In the coding region, the third position of the codon showed high G + C content (86%). The gene was expressed as an unfused protein in Escherichia coli by using an E. coli expression vector. This protein, which reacted with polyclonal antibody raised against alpha antigen from Mycobacterium tuberculosis, would be applicable to the immunodiagnosis of tuberculosis.
Matsuo
K
K
Central Research Laboratories of Ajinomoto Co. Inc., Kawasaki City, Japan.
Yamaguchi
R
R
Yamazaki
A
A
Tasaka
H
H
Yamada
T
T
D000595:Amino Acid Sequence; D000942:Antigens, Bacterial / Q000235:genetics*; D001483:Base Sequence; D003001:Cloning, Molecular; D004262:DNA Restriction Enzymes; D004269:DNA, Bacterial / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics; D005786:Gene Expression Regulation; D005822:Genetic Vectors; D008969:Molecular Sequence Data; D009163:Mycobacterium bovis / Q000235:genetics* / Q000276:immunology; D009693:Nucleic Acid Hybridization; D010957:Plasmids
D000942:Antigens, Bacterial; D004269:DNA, Bacterial; C052242:alpha antigens, Mycobacterium; D004262:DNA Restriction Enzymes