Molecular cloning and expression of Mycobacterium tuberculosis strain Aoyama B peptide antigen genes in Escherichia coli. A gene encoding 15kD antigen (AT 01).:A Gene Encoding 15kD Antigen (AT01)

概要

論文の詳細を見る
A genomic library of Mycobacterium tuberculosis strain Aoyama B in Escherichia coli K-12 was constructed by cloning Sau 3A I cleaved M.tuberculosis Aoyama B chromosomal DNA into pUC 18, pUC 181 or pUC 182.Clones reacting with anti-PPDs-rabbit-serum were screened by immunoblotting among 3×104 clones.Seven clones were selected; designating pAT01, pAT 101, pAT 102, pAT 103, pAT 104, pAT 105 and pAT 201.On Western blotting, they were shown to produce 15kD (pAT 01, pAT 101, pAT 105), 18kD (pAT103) and 60kD (pAT102, pAT201) peptide antigens. Restriction endonuclease map of each of the above clones was composed, and putative coding frames of anti-PPDs-rabbit-serum reactive peptides were deduced by analysing deletion derivatives.Nucleotide sequence of pAT01, encoding for 15kD peptide antigen was analyzed, and a Hinf I-Hinf I fragment in pAT01 was subcloned into pUC 18-E CPL 1to determine the direction of its reading frame.The origin of promoter that drives cloned mycobacterial genes in E.coli was discused.
一般社団法人日本結核病学会の論文