VAAM-Jahrestagung 2012 18.–21. März in Tübingen

161The importance of Leptospirosis is mainly its zoonotic potential, though itcan create losses in domesticated pigs as well. The organism is commonlyfound in bodies of water, moist soil or vegetation contaminated by theurine or tissues of infected animals. For example, swimmers can contractthe disease in contaminated infected waters.In contrast, porcine parvovirosis represents one of the most importantdisorders in domesticated pigs, but is not considered zoonotic. Highporcine parvovirus (PPV) seroprevalences were found in wild boars indifferent European countries and the population of these animals iscurrently increasing. Therefore, wild boars may represent a threat fordomesticated pigs.In the present study, a total of 285 samples of wild boars shot in the areaaround Dresden, Saxony were examined on the presence of antibodiesagainst PPV and Leptospires.The specific antibody titres were determined for PPV byhemagglutination-inhibition test (HI) and for Leptospira spp. bymicroagglutination test (MAT). The MAT panel consisted of 10 serovars.In case of PPV, titres 1:40 and in case of Leptospires, titres 1:100 wereconsidered positive.In total, 54.4 % of the samples were positive for PPV. To our knowledge,this is the first study on PPV seroprevalence in wild boars in Germany andthe results indicate the need for further investigation.Although the examination for leptospirosis has not yet been finished,preliminary results suggest a surprisingly low prevalence of about 2.3 %,with two sera showing MAT-titres of 1:50. In a previous study of urbanwild boars higher seroprevalences (18%) were found.OTP108Non-conventional translation initiation in bacteriaM. Lehr*, P. Ludwig, D.J. Näther, J. SoppaGoethe-University, Institute for Molecular Biosciences, Frankfurt,GermanyInitiation of translation is an important step in the process of geneexpression. As initiation is the rate-limiting step of translation, mostregulatory mechanisms act at this step. The well-studied conventionalpathway of translation initiation in bacteria relies on the interaction of a socalled Shine Dalgarno (SD) motif upstream of the start codon with theanti-SD motif at 3’-end of the 16S rRNA. In addition to conventionaltranscripts two types of non-conventional transcripts exist in bacteria, i.e.leaderless transcripts lacking a 5’-UTR and transcripts with a 5’-UTRwithout a SD motif. Initiation at leaderless transcripts requires thepreassembled 70S ribosome and the initiator tRNA, while the initiationmechanism for transcripts with SD-less 5’-UTRs is still unknown.Only about 60% of all E. coli genes are accompanied by SD motifs andthus about 40% have non-conventional transcripts (1). About 40 geneswith and without SD motifs were chosen and the 5’-ends of theirtranscripts were determined using 5’-RACE. None of the transcripts wasleaderless, while 18 had a 5’-UTR without a SD motif. The 5’-UTRs ofthree SD-less transcripts and of one conventional transcript were fused tothe gusA reporter gene. The initiation efficiencies of two of the SD-lesstranscripts were about 50% compared to the highly expressed conventionalcontrol transcript and even higher during growth at 20 o C, underscoring thatefficient translation is possible in the absence of a SD motif.To gain a genome-wide overview of translational efficiencies in E. colitranslatome analyses were established, i.e. the separation of ribosome-freeuntranslated transcripts and of ribosome-bound transcripts and thecomparison of both fractions using DNA microarrays. As a first approachtranslation under standard conditions was compared to translation in thepresence of Kasugamycin, which was described to specifically inhibittranslation of conventional transcripts. The 5’-ends of selected transcriptswere determined. In contrast to the current belief there was no correlationbetween the presence of an SD motif and the inhibitory effect ofKasugamycin. The next translatome analyses aim at characterizing theinfluence of different stress conditions on translational efficiencies and arecurrently under way.(1) Chang, B, Halgamuge, S., and Tang, S.L. (2006) Analysis of SD sequences in completedmicrobial genomes: non-SD-led genes are as common as SD-led genes. Gene 373: 90-99.OTP109Two new carotenoid cleavage oxygenases from marine bacteriaJ. Hoffmann* 1 , J. Altenbuchner 1 , H. Beuttler 2 , J. Bóna-Lovász 31 University of Stuttgart, Institute of Industrial Genetics, Stuttgart, Germany2 University of Stuttgart, Institute of Technical Biochemistry, Stuttgart, Germany3 University of Stuttgart, Institute for System Dynamics, Stuttgart, GermanyApocarotenoids are carotenoid cleavage products that are predominantlyproduced by carotenoid cleavage oxygenases, a class of non-heme ironenzymes that specifically cleave C-C double bonds of carotenoids.Apocarotenoids have natural functions as colorants, antioxidants, aromacompounds or hormone-like signaling molecules. They are technicallyapplied as nutritional supplements and colorants or flavors for food,cosmetics and pharmaceutical products. To date over 2000 genomes ofeukaryotes, archaea and bacteria have been sequenced and the data werepublished in the GenBank [1]. Numerous of these genomes containputative carotenoid cleavage oxygenase genes that have not beeninvestigated yet. We constructed a two-plasmid expression system fortesting the carotenoid cleavage activities of such enzymes. Two carotenoidcleavage oxygenases from Sphingopyxis alaskensis RB2256 andPlesiocystis pacifica SIR-1 were further investigated.1. D.A. Benson, I. Karsch-Mizrachi, D.J. Lipman, J. Ostell and D.L. Wheeler, Nucleic AcidsResearch 36 (2008), p. D25.OTP110Isotopic fingerprints of bacterial chemosymbiosis in thebivalve Loripes lacteusA. Dreier* 1,2 , L. Stannek 1 , M. Blumenberg 2 , M. Taviani 3 , M. Sigovini 4 ,C. Wrede 1 , V. Thiel 2 , M. Hoppert 1,21 Universität Göttingen, Institut f. Mikrobiologie und Genetik, Göttingen,Germany2 Universität Göttingen, Courant Centre Geobiology, Göttingen, Germany3 ISMAR-CNR, Bologna, Italy4 ISMAR-CNR, Venice, ItalyMetazoans with chemosynthetic bacterial endosymbionts are widespreadin marine habitats and respective endosymbioses are known from sevenrecent animal phyla. However, little is known about endosymbioses infossil settings and, hence, its ecological significance in earth history. In thepresented project, we investigate the ancient and recent bivalve faunaliving at marine sedimentary oxic/anoxic interfaces. Two bivalve speciescollected from the same benthic environment - a Mediterranean lagoon -were studied in detail. The diet of Loripes lacteus is based on thiotrophicgill symbionts whereas Venerupis aureus is a filter feeding bivalve withoutsymbionts. The presence of three key enzymes from sulfur oxidation(APS-reductase), carbon fixation (RubisCO) and assimilation of nitrogen(glutamine synthetase [GS]) were detected by immunofluorescence insymbionts of Loripes and/or by activity tests in living specimens.In search of biosignatures associated with thiotrophic chemosymbionts thatmight be suitable for detection of chemosymbiontic diets in recent andfossil bivalve shells, we analyzed the isotopic composition of shell lipids( 13 C) and the bulk organic matrix of the shell ( 13 C, 15 N, 34 S). We couldshow that the combined 15 N and 13 C values from shell extracts are stablein subfossil (Pleistocene) bivalve specimens, as long as the isotopic data is"calibrated" with respective signatures from a filter feeding bivalvesampled from the same site or lithostratigraphic bed.OTP111A plasmid toolkit for the analysis of regulatory elements inBacillus licheniformisR. Hertel*, H. LiesegangInstitute of Microbiology and Genetics, Göttingen Genomics Laboratory,Göttingen, GermanyBacillus licheniformis is a valuable industrial microorganism. Strains of itsspecies are used in industrial production of enzymes and antibiotics (1). Toimprove the industrial potential of this organism the investigation of theregulation, especially concerning transcriptional and translational features,is of great interest. However, the research on physiological activitiesduring fermentation processes suffers from the lack of adequate moleculartools. Here we present a set of related E. coli-Bacillus shuttle vectors forthe in vivo investigation of promoters and riboswitches. The backbone ofour vectors is constructed in a modular way to ease the adaptation of thedifferent components to a variety of experimental settings. The qualitativeanalysis of single cells in a culture is supported by gfp-vectors. Thequantification of regulatory activities is the target application of our lacZvectors.The origin of replication of the pUB110 (2) vector offers theapplication in other species of the genus Bacillus. The observation that anumber of strains are genetically difficult to access is addressed by theability of the vectors to be transferred by transconjugation. We have shownthat our plasmids can be transferred from E. coli S17-1 (3) as a donorstrain to a number of test strains.(1) Schallmey, M., A. Singh, and O.P. Ward, Can J Microbiol, 2004. 50(1): p. 1-17(2) Keggins, K. M., P. S.Lovett, and E J Duvall, Proc Natl Acad Sci U S A. 1978 March; 75(3): 1423-1427.(3) Simon, R., U. Priefer, and A. Pühler., Bio-Technology, 1983. 9(1): p. 784-791OTP112Characterisation of the cambialistic quercetinase fromStreptomyces sp. FLAD. Nianios*, S. FetznerWestfälische Wilhelms-Universität Münster, Institut für MolekulareMikrobiologie und Biotechnologie, Münster, GermanyQuercetinases catalyse the 2,4-dioxygenolytic cleavage of quercetin(3,5,7,3,4-pentahydroxyflavone), a wide-spread plant flavonol. Theybelong to the cupin superfamily, which is characterised by a six-stranded-barrel fold and conserved amino acid motifs that provide the 3-His-1-Glu ligands to a divalent metal ion [1] . Whereas many cupin-typedioxygenases use Fe 2+ for catalysis, quercetinase (QueD) fromBIOspektrum | Tagungsband 2012