Expression of Mycoplasma pulmonis antigens in Escherichia coli.
The expression of Mycoplasma pulmonis antigen in Escherichia coli was investigated by cloning genomic DNA derived from M. pulmonis m 53, and the DNA fragment participating in antigen expression was identified. When the DNA library of M. pulmonis was screened by colony immunoassay using anti-M. pulmonis serum, 10 recombinant clones expressing seroreactive antigens were obtained. The recombinant plasmids isolated from these clones included 3.7-6.5 kilobase pair (kbp) DNA inserts, while all clones contained a common 2.3-kbp DNA fragment. Subcloning of initial DNA inserts showed that the common 2.3-kbp fragment is essential for antigen expression. Moreover, antiserum against the recombinant antigen generated from the 2.3-kbp DNA fragment recognized a native M. pulmonis antigen. The reactivity of this antiserum was absorbed specifically with M. pulmonis. These results suggest that the cloned 2.3-kbp DNA fragment codes an antigen specific to M. pulmonis.
Kunita
S
S
Department of Laboratory Animal Science, School of Hygienic Science, Kitasato University, Sagamihara, Japan.
Terada
E
E
Ghoda
A
A
Sakurai
Y
Y
Suzuki
H
H
Takakura
A
A
Kagiyama
N
N
D000942:Antigens, Bacterial / Q000235:genetics*; D003001:Cloning, Molecular; D004269:DNA, Bacterial / Q000235:genetics*; D004926:Escherichia coli / Q000235:genetics* / Q000276:immunology; D005815:Genetic Code; D015698:Genomic Library; D009177:Mycoplasma pneumoniae / Q000235:genetics* / Q000276:immunology; D010957:Plasmids
D000942:Antigens, Bacterial; D004269:DNA, Bacterial