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  • 1
    ISSN: 1574-6968
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: Abstract Plasmid pCIT264 from Lactococcus lactis subsp. lactis biovar diacetylactis (L. diacetylactis) contains an insertion sequence (IS)-like element located in the citrate utilization (citQRP) cluster. This 967-nucleotide long element is bounded by 17 bp perfect inverted repeats and contains an open reading frame (ORF1) composed of 296 codons, which could encode a transposase. Expression of the IS from pCIT264 generates two mRNAs of 2900 and 1900 nucleotides. The transcription is driven by the P3 promoter, composed of a — 10 region located at the right end of the IS and of a — 35 region positioned downstream of this element. The IS-like element (IS982) is present in seven copies in the L. diacetylactis genome. The copy present in pCIT264 is highly stable and does not promote rearrangements of the cit cluster. We suggest that the stable maintenance of the IS-like element in pCIT264 could be due to a translational control of the putative transposase by an antisense RNA.
    Type of Medium: Electronic Resource
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  • 2
    ISSN: 1574-6968
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: Abstract The Lactococcus lactis subsp. lactis biovar diacetylactis citrate permease P is encoded by the citP gene borne by an 8.3-kb Cit+ plasmid (F. Sesma, Gardiol, D., de Ruiz Holgado, A.P. and de Mendoza, D., Appl. Environ. Microbiol. 56 (1990) 2099–2103). By subcloning different DNA fragments from the Cit+ plasmid it was found that citP is the only plasmidic gene required for the transport of citrate. Northern blot analysis revealed that citP is transcribed in two mRNA species with lengths of 2.9 and 1.9 kb. The transcriptional pattern, the total amount of citP mRNAs and the transport of citrate were unaffected by the presence of citrate in the growth media. Our results support the conclusion that transcription of citP as well as citrate transport are not induced by citrate in Lactococcus lactis subsp. lactis biovar diacetylactis.
    Type of Medium: Electronic Resource
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  • 3
    ISSN: 1574-6968
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: The citMCDEFGRP cluster from Leuconostoc paramesenteroides involved in citrate utilization was cloned and its nucleotide sequence determined. Homology of the inferred gene products with characterized enzymes reveals that citP encodes the citrate permease P, citC the citrate ligase and citDEF the subunits of the citrate lyase of Leuconostoc. Moreover, it suggests that citM encodes a Leuconostoc malic enzyme. Analysis of citrate consumption by and citrate lyase activity of Lc. paramesenteroides J1[pCITJ1] showed that its citrate permease and its citrate lyase are induced by the presence of citrate in the growth medium. Southern blot analysis demonstrated that the citMCDEFGRP cluster is located in a plasmid.
    Type of Medium: Electronic Resource
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  • 4
    Electronic Resource
    Electronic Resource
    Oxford, UK : Blackwell Publishing Ltd
    FEMS microbiology letters 142 (1996), S. 0 
    ISSN: 1574-6968
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: Abstract Citrate uptake in Lactococcus lactis ssp. lactis biovar diacetylactis (L. diacetylactis) is catalysed by a secondary citrate carrier, the citrate permease P (CitP). In this report the mechanism of citrate uptake was investigated in resting cells of L. diacetylactis. The results presented reveal that the pH gradient (ΔpH) and the membrane potential (ΔΨ) are driving forces for citrate uptake. Moreover, it seems that CitP activity requires neither Na+ nor Mg2+ cations.
    Type of Medium: Electronic Resource
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  • 5
    ISSN: 1617-4623
    Keywords: Key words Transcriptional activation ; Lactococcuslactis ; Escherichia coli ; Plasmid pCIT264 ; Expression of citP gene
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract  The lactococcal plasmid pCIT264 contains a cluster of three genes (citQ, citR and citP) involved in the transport of citrate in Lactococcus lactis biovar diacetylactis. The cit cluster contains a copy of a newly discovered insertion sequence (IS)-like element located between its promoter P1 and the first gene of the cluster. In this report, we show that this IS-like element can act as a mobile switch for the downstream genes, creating two new transcriptional promoters named P2 and P2′. The P2 promoter is recognized by the lactococcal RNA polymerase in vivo. This is a hybrid promoter composed of a −35 region reading outwards 12 bp from the right end of the IS-like element, and a nucleotide sequence from the recipient plasmid, adjacent to the element, which provides an appropriately spaced −10 region. Transcription of the citQRP cluster from this promoter takes place during the exponential and stationary phases of growth in L. lactis. Promoter P2′ is included in the IS-like element and is the only promoter responsible for expression of citP in E. coli. Thus, it appears that the introduction of this element into pCIT264 allows expression of the citQRP cluster in E. coli, and increases its levels of expression in L. lactis.
    Type of Medium: Electronic Resource
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  • 6
    ISSN: 1617-4623
    Keywords: citP gene ; Citrate permease Lactococcus lactis ; Gene expression ; Transcription
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The transport of citrate in Lactococcus lactis biovar diacetylactis is mediated by the citrate permease P. This polypeptide is encoded by the citP gene carried by plasmid pCIT264. In this report, we characterize the citP transcript, identify a cluster of two genes cotranscribed with citP and describe their post-transcriptional regulation. The transcriptional promoter is located 1500 nucleotides upstream of the citP gene and the transcriptional terminator is positioned next to the 3′-end of this gene. The DNA sequence was determined of the region upstream of the citP gene, including the promoter. Two partially overlapping open reading frames, citQ and citR were identified, which could encode polypeptides of 3.9 and 13 kDa respectively. These two genes, together with citP, constitute the cit cluster. Moreover, an IS-like element located between the cit promoter and the citQ open reading frame was identified. This element includes an open reading frame ORF1, which could encode a 33 kDa polypeptide. A translational fusion between the citP and a cat reporter gene showed that translation of citR and citP is coupled, and regulated by CitR. The cit mRNA was subjected to specific cleavage after addition of rifampicin to the bacterial cultures. We propose that expression of the cit cluster is controlled at the post-transcriptional level by mRNA processing at a putative complex secondary structure and by translational repression mediated by CitR.
    Type of Medium: Electronic Resource
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