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Results for “Genomic sequencing”

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At least 595 records · Page 33Linked to original sources
PubMed2005

Use of the Rhodopseudomonas palustris genome sequence to identify a single amino acid that contributes to the activity of a coenzyme A ligase with chlorinated substrates.

Rhodopseudomonas palustris strain RCB100 degrades 3-chlorobenzoate (3-CBA) anaerobically. We purified from this strain a coenzyme A ligase that is active with 3-CBA and determined its N-terminal amino acid sequence to be identical to that of a cyclohexanecarboxylate-CoA ligase encoded by aliA from the R. palustris strain (CGA009) that has been sequenced. Strain CGA009 differs from strain RCB100 in that it does not use 3-CBA as a sole carbon source. The aliA gene from the 3-CBA degrading strain differed by a single nucleotide from the aliA gene from strain CGA009, causing the substitution of a serine for a threonine at position 208. Both AliA enzymes, purified as His-tagged fusion proteins, had comparable activities with cyclohexanecarboxylate. However, AliA from the 3-CBA degrading strain was 10-fold more active with 3-CBA (kcat/Km of 4.3 x 10(4) M(-1) s(-1)) than the enzyme from the sequenced strain (kcat/Km 0.32 x 10(4) M(-1) s(-1)). The CGA009 enzyme was not sufficiently active with 3-CBA to complement an RCB100 aliA mutant for growth on this compound. Here, whole genome sequence information enabled us to identify a single nucleotide among 5.4 million nucleotides that contributes to the substrate preference of a coenzyme A ligase.

Acyl Coenzyme A↗
PubMed2025

The genome sequence of the solitary wasp, Cerceris ruficornis (Fabricius, 1793).

We present a genome assembly from a female specimen of Cerceris ruficornis (solitary wasp; Arthropoda; Insecta; Hymenoptera; Crabronidae). The genome sequence has a total length of 566.08 megabases, of which 65.35% is scaffolded into 14 chromosomal pseudomolecules. The mitochondrial genome has also been assembled, with a length of 18.07 kilobases. Gene annotation of this assembly on Ensembl identified 11,093 protein-coding genes.

Cerceris ruficornis↗
PubMed2000

Transposon mutagenesis of Mycoplasma gallisepticum by conjugation with enterococcus faecalis and determination of insertion site by direct genomic sequencing.

Few genetic systems for studying mycoplasmas exist, but transposon Tn916 has been shown to transpose into the genomes of some species and can be used as an insertional mutagen. In the current study, the ability of Enterococcus faecalis to serve as a donor for the conjugative transfer of transposon Tn916 into the genome of the avian pathogen Mycoplasma gallisepticum strain PG31 was examined. Transconjugants were obtained at a frequency of > or =6 x 10(-8) per recipient CFU. To determine the transposon insertion site, an oligonucleotide primer corresponding to the 3' end of Tn916 was designed for the purpose of directly sequencing genomic DNA without PCR amplification. Using the direct sequencing approach, Tn916 was shown to insert into any of numerous sites in the M. gallisepticum genome. This is the first report of conjugal transposition of Tn916 into the M. gallisepticum genome. The ability to determine transposon insertion sites in mycoplasmas by genomic sequencing has not been previously described and allows rapid sequence analysis of transposon-generated mutants.

Animals↗
PubMed2026

The genome sequence of the Tufted Button, Acleris cristana (Denis & Schiffermüller, 1775).

We present a genome assembly from an individual female Acleris cristana (the Tufted Button; Arthropoda; Insecta; Lepidoptera; Tortricidae). The genome sequence is 562.6 megabases in span. Most of the assembly is scaffolded into 31 chromosomal pseudomolecules, including the W and Z sex chromosomes. The mitochondrial genome has also been assembled and is 16.1 kilobases in length. Gene annotation of this assembly on Ensembl identified 12,598 protein coding genes.

Acleris cristana↗
PubMed2023

The genome sequence of the Six-striped Rustic, Xestia sexstrigata (Haworth, 1809).

We present a genome assembly from an individual female Xestia sexstrigata (the Six-striped Rustic; Arthropoda; Insecta; Lepidoptera; Noctuidae). The genome sequence is 638.3 megabases in span. Most of the assembly is scaffolded into 32 chromosomal pseudomolecules, including the W and Z sex chromosomes. The mitochondrial genome has also been assembled and is 15.36 kilobases in length. Gene annotation of this assembly on Ensembl identified 15,104 protein coding genes.

Lepidoptera↗
PubMed1999

The malaria genome sequencing project: complete sequence of Plasmodium falciparum chromosome 2.

An international consortium has been formed to sequence the entire genome of the human malaria parasite Plasmodium falciparum. We sequenced chromosome 2 of clone 3D7 using a shotgun sequencing strategy. Chromosome 2 is 947 kb in length, has a base composition of 80.2% A + T, and contains 210 predicted genes. In comparison to the Saccharomyces cerevisiae genome, chromosome 2 has a lower gene density, a greater proportion of genes containing introns, and nearly twice as many proteins containing predicted non-globular domains. A group of putative surface proteins was identified, rifins, which are encoded by a gene family comprising up to 7% of the protein-encoding gene in the genome. The rifins exhibit considerable sequence diversity and may play an important role in antigenic variation. Sixteen genes encoded on chromosome 2 showed signs of a plastid or mitochondrial origin, including several genes involved in fatty acid biosynthesis. Completion of the chromosome 2 sequence demonstrated that the A + T-rich genome of P. falciparum can be sequenced by the shotgun approach. Within 2-3 years, the sequence of almost all P. falciparum genes will have been determined, paving the way for genetic, biochemical, and immunological research aimed at developing new drugs and vaccines against malaria.

Animals↗
PubMed2004

DNAskew: statistical analysis of base compositional asymmetry and prediction of replication boundaries in the genome sequences.

Sueoka and Lobry declared respectively that, in the absence of bias between the two DNA strands for mutation and selection, the base composition within each strand should be A=T and C=G (this state is called Parity Rule type 2, PR2). However, the genome sequences of many bacteria, vertebrates and viruses showed asymmetries in base composition and gene direction. To determine the relationship of base composition skews with replication orientation, gene function, codon usage biases and phylogenetic evolution, in this paper a program called DNAskew was developed for the statistical analysis of strand asymmetry and codon composition bias in the DNA sequence. In addition, the program can also be used to predict the replication boundaries of genome sequences. The method builds on the fact that there are compositional asymmetries between the leading and the lagging strand for replication. DNAskew was written in Perl script language and implemented on the LINUX operating system. It works quickly with annotated or unannotated sequences in GBFF (GenBank flatfile) or fasta format. The source code is freely available for academic use at http://www.epizooty.com/pub/stat/DNAskew.

Algorithms↗
PubMed2006

A second generation radiation hybrid map to aid the assembly of the bovine genome sequence.

BACKGROUND: Several approaches can be used to determine the order of loci on chromosomes and hence develop maps of the genome. However, all mapping approaches are prone to errors either arising from technical deficiencies or lack of statistical support to distinguish between alternative orders of loci. The accuracy of the genome maps could be improved, in principle, if information from different sources was combined to produce integrated maps. The publicly available bovine genomic sequence assembly with 6x coverage (Btau_2.0) is based on whole genome shotgun sequence data and limited mapping data however, it is recognised that this assembly is a draft that contains errors. Correcting the sequence assembly requires extensive additional mapping information to improve the reliability of the ordering of sequence scaffolds on chromosomes. The radiation hybrid (RH) map described here has been contributed to the international sequencing project to aid this process. RESULTS: An RH map for the 30 bovine chromosomes is presented. The map was built using the Roslin 3000-rad RH panel (BovGen RH map) and contains 3966 markers including 2473 new loci in addition to 262 amplified fragment-length polymorphisms (AFLP) and 1231 markers previously published with the first generation RH map. Sequences of the mapped loci were aligned with published bovine genome maps to identify inconsistencies. In addition to differences in the order of loci, several cases were observed where the chromosomal assignment of loci differed between maps. All the chromosome maps were aligned with the current 6x bovine assembly (Btau_2.0) and 2898 loci were unambiguously located in the bovine sequence. The order of loci on the RH map for BTA 5, 7, 16, 22, 25 and 29 differed substantially from the assembled bovine sequence. From the 2898 loci unambiguously identified in the bovine sequence assembly, 131 mapped to different chromosomes in the BovGen RH map. CONCLUSION: Alignment of the BovGen RH map with other published RH and genetic maps showed higher consistency in marker order and chromosome assignment than with the current 6x sequence assembly. This suggests that the bovine sequence assembly could be significantly improved by incorporating additional independent mapping information.

Animals↗
PubMed2006

Phytophthora genome sequences uncover evolutionary origins and mechanisms of pathogenesis.

Draft genome sequences have been determined for the soybean pathogen Phytophthora sojae and the sudden oak death pathogen Phytophthora ramorum. Oömycetes such as these Phytophthora species share the kingdom Stramenopila with photosynthetic algae such as diatoms, and the presence of many Phytophthora genes of probable phototroph origin supports a photosynthetic ancestry for the stramenopiles. Comparison of the two species' genomes reveals a rapid expansion and diversification of many protein families associated with plant infection such as hydrolases, ABC transporters, protein toxins, proteinase inhibitors, and, in particular, a superfamily of 700 proteins with similarity to known oömycete avirulence genes.

Algal Proteins↗
PubMed2022

The genome sequence of the plain-faced dronefly, Eristalis arbustorum (Linnaeus, 1758).

We present a genome assembly from an individual female Eristalis arbustorum (the plain-faced dronefly; Arthropoda; Insecta; Diptera; Syriphidae). The genome sequence is 451 megabases in span. The majority of the assembly (94.71%) is scaffolded into 6 chromosomal pseudomolecules, with the X sex chromosome assembled. The complete mitochondrial genome was also assembled and is 16.0 kilobases in length.

Diptera↗
PubMed2004

Transition metal transport in the green microalga Chlamydomonas reinhardtii--genomic sequence analysis.

Uptake and export systems play a major role in transition metal homeostasis. The objective of this study was to identify potential metal transport mechanisms in the green microalga Chlamydomonas reinhardtii. We concentrated on the four major transition metal transporter families found in plants and other organisms: the ZIP, CDF and Nramp families, and the CPx-ATPases. Using the information available for these protein families we performed comparative sequence analysis in the recently released genome of C. reinhardtii. Using this approach we were able to identify members of all four transporter families (four ZIPs, one CDF, two CPx-ATPases, and five Nramps). These findings advance our current knowledge of the metal transport processes present in C. reinhardtii. In addition, by subsequent in silico splicing of the genomic sequence we obtained cDNA sequences which led to the identification of ESTs (expressed sequence tags) in the C. reinhardtii EST database. These identified ESTs will be valuable for the cloning and characterization of several metal transporters utilized by the alga.

Amino Acid Sequence↗
PubMed2006

Leeuwenhoekiella blandensis sp. nov., a genome-sequenced marine member of the family Flavobacteriaceae.

Bacteria in the family Flavobacteriaceae are increasingly recognized to play important roles in the degradation of organic matter during and following algal blooms. A novel heterotrophic, rod-shaped, aerobic, yellow-pigmented and gliding bacterium was isolated from a seawater sample collected in the Bay of Blanes in the north-western Mediterranean Sea. Analysis of its 16S rRNA gene sequence, retrieved from the whole-genome sequence, showed that the bacterium was closely related to members of the genus Leeuwenhoekiella within the family Flavobacteriaceae, phylum Bacteroidetes. Phenotypic, genotypic, chemotaxonomic and phylogenetic analyses supported the creation of a novel species to accommodate this bacterium, for which the name Leeuwenhoekiella blandensis sp. nov. is proposed. The type strain is MED 217(T) (=CECT 7118(T)=CCUG 51940(T)).

Anti-Bacterial Agents↗
PubMed2025

Identifying healthcare transmission routes of nontuberculous mycobacteria with whole genome sequencing: a systematic review.

OBJECTIVE: To enumerate and describe the effect of whole genome sequencing (WGS) on epidemiological investigations of healthcare-associated transmission of nontuberculous mycobacteria (NTM). DESIGN: Systematic review. METHODS: We performed a literature search using targeted search terms to identify articles meeting inclusion criteria. Data extraction of study characteristics and outcomes was performed by two independent researchers. The primary outcome was the author interpretation of WGS utility in the investigation of suspected healthcare-associated transmission of NTM. The secondary outcome was whether a transmission route was identified through WGS. RESULTS: Thirty-one studies were included in the final analysis with 28 (90%) concluding that WGS was helpful in transmission investigations and in 19 of these 28 (68%) WGS aided in identifying a transmission route. The most common identified transmission routes were water-borne point sources (10), heater-cooler units (6), patient-to-patient (4), and a healthcare worker (1). CONCLUSION: WGS is an informative tool in investigating healthcare transmission of NTM.

Humans↗
PubMed2024

The genome sequence of the English holly, Ilex aquifolium L. (Aquifoliaceae).

We present a genome assembly from an individual Ilex aquifolium (the English holly; Eudicot; Magnoliopsida; Aquifoliales; Aquifoliaceae). The genome sequence is 800.0 megabases in span. Most of the assembly is scaffolded into 20 chromosomal pseudomolecules. The assembled mitochondrial and plastid genomes have lengths of 538.43 kilobases and 157.52 kilobases in length, respectively.

Aquifoliales↗
PubMed2006

Full-length genomic sequencing and analysis of four HIV type 1 subtype B isolates circulating in the territory of Russia.

We describe four full-length genomic sequences of HIV-1 subtype B isolates from Russia. These full-length HIV-1 genomes were amplified by nested polymerase chain reaction and sequenced. The sequences obtained were subjected to neighbor-joining phylogenetic analysis using a Kimura two-parameter model and to detailed sequence analysis. Comparison of the sequences obtained with 68 near full-length HIV-1 subtype B sequences from different geographic regions of the world revealed that isolates from Russia did not cluster significantly with other subtype B genomes. Sequences AY819715 and AY751407 significantly formed one cluster, which indicates their close similarity. The HIV-1 genomes obtained from Russia did not form a distinct homogeneous group inside subtype B. Their genetic diversity probably reflects the result of multiple introduction events of different subtype B strains to Russia. Sequences AY819715 and AY751406 possess some features of viruses from long-term survivors, such as specific extensions of the Env V2 region.

Female↗
PubMed1997

Genomic sequence and organization of the human gene for cytochrome c oxidase subunit (COX7A1) VIIa-M.

Cytochrome c oxidase (COX, EC 1.9.3.1), the last component of the mitochondrial electron transfer chain, is built up by 13 polypeptides; 3 of them are encoded by the mitochondrial genome while the 10 smaller subunits are encoded by the nuclear genome. Several nuclear-encoded subunits occur in two different tissue-specific isoforms, a constitutive "L"-form and an "M"-form specific for skeletal and heart muscle. In this article, we describe the genomic sequence and organization of the human gene for COX subunit VIIa-M (COX7A1) located on chromosome 19q13.1 and compare it to its bovine homologue. The coding region of the gene extends over 1.45 kb of genomic sequence, organized in four exons. Intron-exon boundaries are well conserved between cattle and humans. Although it is a gene for a tissue-specific isoform, it has some features of a housekeeping gene: it is located in a CpG island, like its bovine homologue, and no TATA or CCAAT boxes were found in the 5' flanking sequence. Southern hybridization of COX7A1 to human genomic DNA revealed no pseudogenes. Putative binding sites for ubiquitous transcription factors like Sp1 and specific expression in skeletal as well as in heart muscle have been found. In contrast to the bovine gene, the human gene contains putative binding sites for nuclear respiratory factor 2 (NRF-2), which is implicated in the activation of other respiratory enzymes. Therefore, the human and the bovine genes, although well conserved in their coding regions, could differ in the tissue-specific regulation of gene expression. Knowledge of the gene structure will facilitate the analysis of the involvement of subunit VIIa in mitochondrial myopathies and may provide clues to the function of this subunit in a multicomponent enzyme.

Animals↗