Nucleotide sequence and expression in Escherichia coli of the Lactococcus lactis citrate permease gene.

Abstract:

:The plasmid-encoded citrate determinant of the Lactococcus lactis subsp. lactis var. diacetylactis NCDO176 was cloned and functionally expressed in a Cit- Escherichia coli K-12 strain. From deletion derivative analysis, a 3.4-kilobase region was identified which encodes the ability to transport citrate. Analysis of proteins encoded by the cloned fragment in a T7 expression system revealed a 32,000-dalton protein band, which correlated with the ability of cells to transport citrate. Energy-dependent [1,5-14C]citrate transport was found with membrane vesicles prepared from E. coli cells harboring the citrate permease-expressing plasmid. The gene encoding citrate transport activity, citP, was located on the cloned fragment by introducing a site-specific mutation that abolished citrate transport and resulted in a truncated form of the 32,000-dalton expression product. The nucleotide sequence for a 2.2-kilobase fragment that includes the citP gene contained an open reading frame of 1,325 base pairs coding for a very hydrophobic protein of 442 amino acids, which shows no sequence homology with known citrate carriers.

journal_name

J Bacteriol

journal_title

Journal of bacteriology

authors

David S,van der Rest ME,Driessen AJ,Simons G,de Vos WM

doi

10.1128/jb.172.10.5789-5794.1990

subject

Has Abstract

pub_date

1990-10-01 00:00:00

pages

5789-94

issue

10

eissn

0021-9193

issn

1098-5530

journal_volume

172

pub_type

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