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1Microbial Ingredients Section, NlZO Food Research, PO Box 20, 6710 BA Ede, The Netherlands
2Laboratory for Food Microbiology, Food Science Group, Wageningen University and Research Center, Wageningen, The Netherlands
3Department of Genetics, Groningen University, Haren, The Netherlands
1 Author for correspondence: Oscar P. Kuipers. Tel: +31 318 659525. Fax: +31 318 650400.e-mail: kuipers@nizo.nl
ABSTRACT
SUMMARY: A family of genes encoding cold-shock proteins, named cspA, cspB, cspC, cspD and cspE, was cloned and sequenced from Lactococcus lactis MG1363. The genes cspA and cspB and the genes cspC and cspD are located in tandem repeats, an organization of csp genes that has never been encountered before. The five genes encode small (7.1-706 kDa) proteins with high mutual sequence identities (up to 85 O/O) and high identities (about 45-65 %) with the major cold- shock proteins from Escherichia coli (CspA) and Bacillus subtilis (CspB)., Northern-blot analysis revealed single transcripts of about 300 nucleotides for each csp gene and showed that cspA, cspB, cspC and cspD mRNA levels were strongly increased upon cold shock to 10 "C (about lo-, 40-, 10- and 30-fold compared to 30 "C, respectively), whereas the cspE mRNA level was not increased. The expression of the cold-induced csp genes was highest in the. 6-8 h lag phase after cold shock. A differential expression in time, in which cspA and cspC were maximally expressed at 2 h and cspB and cspD at 4 h after cold shock, was observed. The -35 and -10 regions of the five promoters were identified and transcriptional start sites were mapped in each case by primer extension at different temperatures which confirmed that regulation takes place at the transcriptional level. Significant differences were observed between the 5'-untranslated leader regions of the four cold-induced csp genes and the corresponding region of the non-cold-induced cspE gene.
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