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Recent genetic studies have elucidated that carbonic anhydrase (CA; EC 4.2.1.1), a ubiquitous enzyme catalyzing interconversion
between CO2 and bicarbonate, is essential for microbial growth under ambient air but not under high-CO2 air. The irregular distribution of the phylogenetically distinct types of CA in the prokaryotic genome suggests its complex
evolutionary history in prokaryotes. This paper deals with the genetic defect of CA in Symbiobacterium thermophilum, a syntrophic bacterium that effectively grows on CO2 generated by other bacteria. Phylogenetic analysis based on 31 ribosomal protein sequences demonstrated the affiliation of
Symbiobacterium with the class Clostridia with 100% bootstrap support. The phylogeny of β- and γ-type CA distributed among Clostridia supported
the view that S. thermophilum and several related organisms lost this enzyme during the course of evolution. The loss of CA could be based on the availability
of a high level of CO2 in their living environments.
Electronic supplementary material The online version of this article (doi:) contains supplementary material, which is available to authorized users. 相似文献
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Arabidopsis ACT2 represents an ancient class of vegetative plant actins and is strongly and constitutively expressed in almost all Arabidopsis sporophyte vegetative tissues. Using the beta glucuronidase report system, the studies showed that ACT2 5′ regulatory region was significantly more active than CaMV 35S promoter in Arabidopsis seedlings and gametophyte vegetative tissues of Physcomitrella patens. Its activity was also observed in rice and maize seedlings. Thus, the ACT2 5′ regulatory region could potentially serve as a strong regulator to express a transgene in divergent plant species. ACT2 5′ regulatory region contained 15 conserved sequence elements, an ancient intron in its 5′ un-translated region (5′ UTR),
and a purine-rich stretch followed by a pyrimidine-rich stretch (PuPy). Mutagenesis and deletion analysis illustrated that
some of the conserved sequence elements and the region containing PuPy sequences played regulatory roles in Arabidopsis. Interestingly, mutation of the conserved elements did not lead a dramatic change in the activity of ACT2 5′ regulatory region. The ancient intron in ACT2 5′ UTR was required for its strong expression in both Arabidopsis and P. patens, but did not fully function as a canonical intron. Thus, it was likely that some of the conserved sequence elements and gene
structures had been preserved in ACT2 5′ regulatory region over the course of land plant evolution partly due to their functional importance. The studies provided
additional evidences that identification of evolutionarily conserved features in non-coding region might be used as an efficient
strategy to predict gene regulatory elements. 相似文献
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The intron sequence of chloroplast rpS16 and the secondary structure of its pre-mRNA were characterized for the first time in 26 Allium sativum accessions of different ecologo-geographical origins and seven related Allium species. The boundaries and main stem-loop consensus sequences were identified for all six domains of the intron. Polymorphism
was estimated for the total intron and its regions. The structural regions of the rpS16 intron proved to be heterogeneous for mutation rate and spectrum. Mutations were most abundant in domains II and IV, and
transition predominated in domains I, III, V, and VI. In addition to structural elements and motifs typical for group IIB
introns, several Allium-specific micro- and macrostructural mutations were revealed. A 290-bp deletion involving domains III and IV and part of domain
V was observed in A. altaicum, A. fistulosum, and A. schoenoprasum. Several indels and nucleotide substitutions were found to cause a deviation of the pre-mRNA secondary structure from the
consensus model of group II introns. 相似文献
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L. Wanntorp H.-E. Wanntorp B. Oxelman M. Källersjö 《Plant Systematics and Evolution》2001,226(1-2):85-107
The phylogeny of the genus Gunnera is investigated for the first time. Twelve species representing the six currently recognised subgenera are analysed. Two
chloroplast DNA regions, the rbcL gene and the rps16 intron, together provide 46 informative characters out of 2335. A combined analysis of both genes gives four most parsimonious
trees, firmly establishing the east South American G. herteri as sister group to the rest of the genus. The African G. perpensa is sister group to two well-supported clades, one including the South American subgenera Misandra and Panke, the other the Australian/New Zealand/Malayan species of subgenera Milligania and Pseudogunnera. Thus, South America is a composite area for Gunnera, showing up at two different levels in the cladogram. Our analysis supports a close biogeographic relationship between Australia
and New Zealand. The evolution of some morphological characters is discussed. Lastly, the unusual structure of some of the
rbcL sequences is reported.
Received July 6, 2000 Accepted October 24, 2000 相似文献
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Krzysztof Kur Iwona Gabriel Joachim Morschhäuser Francesco Barchiesi Elisabetta Spreghini Sławomir Milewski 《Mycopathologia》2010,170(6):397-402
Two genes, LYS21 and LYS22, encoding isoforms of homocitrate synthase, an enzyme catalysing the first committed step in the lysine biosynthetic pathway,
were disrupted in Candida albicans using the SAT1 flipper strategy. The double null lys21Δ/lys22Δ mutant lacked homocitrate synthase activity and exhibited lysine auxotrophy in minimal media that could be fully rescued
by the addition of 0.5–0.6 mM l-lysine. On the other hand, its virulence in vivo in the model of disseminated murine candidiasis appeared identical to that
of the mother, wild-type strain. These findings strongly question a possibility of exploitation of homocitrate synthase and
possibly also other enzymes of the lysine biosynthetic pathway as targets in chemotherapy of disseminated fungal infections. 相似文献
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Hiroyoshi Kubo 《Mycoscience》2009,50(5):400-406
Pilobolus crystallinus shows unique photoresponses at various growing stages. cDNAs for putative photoreceptors were cloned from this fungus. Three
genes named pcmada1, pcmada2, and pcmada3 were identified from the PCR fragments, and amplified with degenerated primers for the LOV domain, which is conserved in
many blue-light receptors. Deduced amino acid sequences for PCMADA1, PCMADA2, and PCMADA3 had one light-oxygen-voltage (LOV)-sensing
and two PER-ARNT-SIM (PAS) domains. A zinc finger DNA-binding motif was conserved in the C-terminals of PCMADA1 and PCMADA3.
However, PCMADA2 lacked the zinc finger motif. Expression of pcmada1 was suppressed by blue light whereas that of pcmada3 was promoted by blue-light irradiation. 相似文献
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Megan N. Hemming Sarah Fieg W. James Peacock Elizabeth S. Dennis Ben Trevaskis 《Molecular genetics and genomics : MGG》2009,282(2):107-117
Activity of the VERNALIZATION1 (VRN1) gene is required for flowering in temperate cereals such as wheat and barley. In varieties that require prolonged exposure
to cold to flower (vernalization), VRN1 is expressed at low levels and is induced by vernalization to trigger flowering. In other varieties, deletions or insertions
in the first intron of the VRN1 gene are associated with increased VRN1 expression in the absence of cold treatment, reducing or eliminating the requirement for vernalization. To characterize natural
variation in VRN1, the first intron of the barley (Hordeum vulgare) VRN1 gene (HvVRN1) was assayed for deletions or insertions in a collection of 1,000 barleys from diverse geographical regions. Ten alleles of
HvVRN1 containing deletions or insertions in the first intron were identified, including three alleles that have not been described
previously. Different HvVRN1 alleles were associated with differing levels of HvVRN1 expression in non-vernalized plants and with different flowering behaviour. Using overlapping deletions, we delineated regions
in the HvVRN1 first intron that are associated with low levels of HvVRN1 expression in non-vernalized plants. Deletion of these intronic regions does not prevent induction of HvVRN1 by cold or the maintenance of increased HvVRN1 expression following cold treatment. We suggest that regions within the first intron of HvVRN1 are required to maintain low levels of HvVRN1 expression prior to winter but act independently of the regulatory mechanisms that mediate induction of HvVRN1 by cold during winter.
Electronic supplementary material The online version of this article (doi:) contains supplementary material, which is available to authorized users.
Nucleotide sequence data reported are available in the DDBJ/EMBL/GenBank databases under the accession numbers 1179825, 1179833,
1179836, 1179858. 相似文献
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Khomaini Hasan Wangsa Tirta Ismaya Idar Kardi Yandi Andiyana Susanti Kusumawidjaya Safri Ishmayana Toto Subroto Soetijoso Soemitro 《Biologia》2008,63(6):1044-1050
α-Amylase from Saccharomycopsis fibuligera R-64 was successfully purified by butyl Toyopearl hydrophobic interaction chromatography, followed by Sephadex G-25 size
exclusion and DEAE Toyopearl anion exchange chromatography. The enzyme has a molecular mass of 54 kDa, as judged by SDS PAGE
analysis. Upon tryptic digestion, two major fragments with relative molecular masses of 39 kDa and 10 kDa, which resemble
the A/B and C-terminal domains in the homologous Taka-amylase, were obtained and were successfully separated with the Sephadex
G-50 size exclusion column. The 39-kDa fragment demonstrated a similar amylolytic activity to that of the undigested enzyme.
However, it was found that the K
m value of the 39-kDa fragment was about two-times higher than that of the undigested enzyme. Moreover, thermostability studies
showed a lower half-life time for the 39-kDa fragment. These findings suggest that the 39-kDa fragment is the catalytic domain,
while the 10-kDa fragment is the C-terminal one, which plays a role in thermostability and starch binding. Although the undigested
enzyme is able to act on raw starches at room temperature, with maize starches as the best substrate, neither the undigested
enzyme nor the fragments adsorb the tested raw starches. These results propose Saccharomycopsis fibuligera α-amylase as a raw starch-digesting but not adsorbing amylase, with a similar domain organization to that of Taka-amylase A. 相似文献
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Wang Minqin Zhao Junsheng Peng Zhenying Guo Wei Wang Yun Wang Le Xia Guangmin 《Plant Cell, Tissue and Organ Culture》2008,92(2):121-130
In order to investigate chromosome elimination in symmetric somatic hybridization between Bupleurum scorzonerifolium and Arabidopsis thaliana, protoplasts were isolated from suspension cultures of both A. thaliana and B. scorzonerifolium parents. Biparental protoplasts were mixed at a rate of 1.5:1 and fused with PEG-method. After protoplast fusion, the products
were cultured in the P5 liquid medium for microcallus formation. Single cell lines formed from microcalli after subculturing
on the MB1 (Xia and Chen, Plant Sci 120:197–203, 1996) solid medium. The putative somatic hybrid cell lines were identified
by cytological and molecular analysis. Of the 132 somatic cell lines generated, 16 were identified as somatic hybrids, with
the phenotypes resembled B. scorzonerifolium parent. These hybrids showed a complete set of B. scorzonerifolium chromosome and 0–2 small chromosome(s) of A. thaliana. A few of them showed nuclear and cytoplasmic SSR fragments of A. thaliana. These hybrid cell lines could differentiate to green spots, buds/leaves through complementation of regeneration ability.
The chromosomes elimination of A. thaliana was discussed.
Wang Minqin and Zhao Junsheng contributed equally to the work. 相似文献
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Amy S. Gargis Anna-Lee D. O’Rourke Gary L. Sloan Robin S. Simmonds 《Journal of molecular evolution》2009,68(5):498-505
Zoocin A is a streptococcolytic enzyme produced by Streptococcus equi subsp. zooepidemicus strain 4881. The zoocin A gene (zooA) and the gene specifying resistance to zoocin A (zif) are adjacent on the chromosome and are divergently transcribed. Twenty-four S. equi subsp. zooepidemicus strains were analyzed to determine the genetic difference among three previously characterized as zoocin A producers (strains
4881, 9g, and 9h) and the 21 nonproducers. LT-PCR and Southern hybridization studies revealed that none of the nonproducer
strains possessed zooA or zif. RAPD and PFGE showed that the 24 strains were a genetically diverse population with eight RAPD profiles. S. equi subsp. zooepidemicus strains 9g and 9h appeared to be genetically identical to each other but quite different from strain 4881. Sequences derived
from 4881 and 9g showed that zooA and zif were integrated into the chromosome adjacent to the gene flaR. A comparison of these sequences with the genome sequences of S. equi subsp. zooepidemicus strains H70 and MGCS10565 and S. equi subsp. equi strain 4047 suggests that flaR flanks a region of genome plasticity in this species.
Electronic supplementary material The online version of this article (doi:) contains supplementary material, which is available to authorized users. 相似文献
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Eric P. Bennett Ya-Wen Chen Tilo Schwientek Ulla Mandel Katrine ter-Borch Gram Schjoldager Stephen M. Cohen Henrik Clausen 《Glycoconjugate journal》2010,27(4):435-444
The Drosophila l(2)35Aa gene encodes a UDP-N-acetylgalactosamine: Polypeptide N-acetylgalactosaminyltransferase, essential for embryogenesis and development (J. Biol. Chem. 277, 22623–22638; J. Biol. Chem. 277, 22616–22). l(2)35Aa, also known as pgant35A, is a member of a large evolutionarily conserved family of genes encoding polypeptide GalNAc-transferases. Phylogenetic and
functional analyses have proposed that subfamilies of orthologous GalNAc-transferase genes are conserved in species, suggesting
that they serve distinct functions in vivo. Based on sequence alignments, pgant35A and human GALNT11 are thought to belong to a distinct subfamily. Recent in vitro studies have shown that pgant35A and pgant7, encoding enzymes from different subfamilies, prefer different acceptor substrates, whereas the orthologous pgant35A and human GALNT11 gene products possess, 1) conserved substrate preferences and 2) similar acceptor site preferences in vitro. In line with the in vitro pgant7 studies, we show that l(2)35Aa lethality is not rescued by ectopic pgant7 expression. Remarkably and in contrast to this observation, the human pgant35A ortholog, GALNT11, was shown not to support rescue of the l(2)35Aa lethality. By use of genetic “domain swapping” experiments we demonstrate, that lack of rescue was not caused by inappropriate
sub-cellular targeting of functionally active GalNAc-T11. Collectively our results show, that fly embryogenesis specifically
requires functional pgant35A, and that the presence of this gene product during fly embryogenesis is functionally distinct from other Drosophila GalNAc-transferase isoforms and from the proposed human ortholog GALNT11. 相似文献
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Gwénaelle Crénès Corinne Moundras Marie-Véronique Demattei Yves Bigot Agnès Petit Sylvaine Renault 《Genetica》2010,138(5):509-517
The eukaryotic transposon Mos1 is a class-II transposable element that moves using a “cut-and-paste” mechanism in which the transposase is the only protein
factor required. The formation of the excision complex is well documented, but the integration step has so far received less
investigation. Like all mariner-like elements, Mos1 was thought to integrate into a TA dinucleotide without displaying any other target selection preferences. We set out to
synthesize what is currently known about Mos1 insertion sites, and to define the characteristics of Mos1 insertion sequences in vitro and in vivo. Statistical analysis can be used to identify the TA dinucleotides that are non-randomly
targeted for transposon integration. In vitro, no specific feature determining target choice other than the requirement for
a TA dinucleotide has been identified. In vivo, data were obtained from two previously reported integration hotspots: the
bacterial cat gene and the Caenorhabditis elegans rDNA locus. Analysis of these insertion sites revealed a preference for TA dinucleotides that are included in TATA or TA × TA
motifs, or located within AT-rich regions. Analysis of the physical properties of sequences obtained in vitro and in vivo
do not help to explain Mos1 integration preferences, suggesting that other characteristics must be involved in Mos1 target choice. 相似文献
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Summary. The paper describes two methods of the synthesis of ethyl (3R,4S)- and (3S,4S)-4-[(benzyloxycarbonyl)amino]-5-[(tert-butyloxycarbonyl)amino]-3-hydroxypentanoates, useful for the syntheses of edeine analogs. Differently N-protected (S)-2,3-diaminopropanoic acid was used as a substrate in both procedures. The absolute configuration of newly generated asymmetric
carbon atoms C-3 in β-hydroxy-γ,δ-diamino products was assigned by means of 1H NMR spectroscopy after their transformation into corresponding piperidin-2-ones.
Received May 24, 2002 Accepted October 10, 2002 Published online December 18, 2002
Acknowledgment The authors are indebted to the Faculty of Chemistry, Technical University of Gdańsk for financial support.
Authors' address: Zbigniew Czajgucki, M. Sc., Department of Pharmaceutical Technology and Biochemistry, Faculty of Chemistry, Technical University
of Gdańsk, 11/12 Narutowicza St., 80-952 Gdańsk, Poland, Fax +48 58 347 11 44, E-mail: zmczaj@wp.pl 相似文献