Open AccessCloning and sequences of inducible and constitutive macrolide resistance genes in Staphylococcus aureus that correspond to an ABC transporter
Author(s) -
Matsuoka Mayumi,
Jánosi László,
Endou Kikutarou,
Nakajima Yoshinori
Publication year - 1999
Publication title -
fems microbiology letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.899
H-Index - 151
eISSN - 1574-6968
pISSN - 0378-1097
DOI - 10.1111/j.1574-6968.1999.tb08830.x
Subject(s) - staphylococcus aureus , atp binding cassette transporter , microbiology and biotechnology , cloning (programming) , gene , biology , genetics , transporter , computational biology , bacteria , computer science , programming language
A restriction map was made and the DNA sequence was determined for a plasmid, pMC38, derived from the inducible macrolide resistance plasmid pEP2104, that showed constitutive resistance to PMS antibiotics (partial macrolide and streptogramin B antibiotics). A 5.04 kb Sal I‐ Pst I fragment (fragment C) of pMC38, which encoded PMS resistance, was cloned into a shuttle vector, pRIT5, to yield pMR504. The transformant Staphylococcus aureus 4220 (pMR504) exhibited constitutive PMS resistance. Fragment C was subcloned to pUC19 in order to determine the DNA sequence. This sequence was consequently found to contain three open reading frames (ORF1–3), of which ORF3 corresponded to the 63 kDa membrane protein (MsrSA) that expressed PMS resistance. According to DNA sequence comparison of the control region of ORF3 in pMC38 and pEP2104, 44 nucleotides including RBS1 and the leader peptide (MTASMRLK) were deleted on plasmid pMC38. This suggests that the leader peptide is essential for the inducible expression of PMS resistance.