Open AccessCloning of chromosomal genes of Lactococcus by heterologous complementation: Partial characterisation of a putative lactose transport gene
Author(s) -
Ross R.,
O'Gara F.,
Condon S.
Publication year - 1989
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.1574-6968.1989.tb03575.x
Subject(s) - cosmid , complementation , lactococcus lactis , biology , plasmid , genetics , gene , subcloning , escherichia coli , genomic library , microbiology and biotechnology , lactococcus , molecular cloning , cloning (programming) , mutant , bacteria , gene expression , peptide sequence , lactic acid , computer science , programming language
A cosmid gene library of the genome of Lactococcus lactis subsp. lactis 712 has been constructed in the broad host range plasmid pLAFR1 in Escherichia coli LE392. Three lactococcal genes from the bank were identified by heterologous complementation of specific mutations in strains of E. coli . A cosmid clone encoding a putative lactose transport gene was identified by complementing an E. coli lac Y mutant. The complemented clone supported the uptake of 14 C lactose in transport assays. The DNA fragment responsible was subcloned and localised to a 1.28 kb fragment of the lactococcal chromosome.