Gene/Protein Disease Symptom Drug Enzyme Compound
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Query: EC:3.1.30.2 (endonuclease)
 18,621 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

Changes in the classification of invertebrate iridoviruses (IVs) (Iridoviridae) have recently been proposed (Williams and Cory, 1994). The previous system of naming isolates according to the host and sequence of discovery (IV type 1, IV2, IV3, etc.) is not adequate for the purposes of taxonomy, since iridovirus isolates may infect many species, including hosts from diverse invertebrate orders. The new system of invertebrate iridovirus nomenclature, as with several other virus families, is based on geographical origin. Proposals have been made, based on DNA hybridization and other characteristics, by which invertebrate iridovirus isolates can be assigned to one of four recognized complexes, or considered as candidates for alternative assignations. This study reports comparative data on the DNA of 14 invertebrate iridovirus isolates used in the Williams and Cory study plus the two type vertebrate iridoviruses, frog virus 3 and flounder lymphocystis disease virus. DNA studies support the validity of assigning several isolates a common name and of grouping the known isolates into four complexes. The detection of such complexes is in broad agreement with previous serological studies. A previously undescribed isolate (San Miguel IV) obtained from the lepidopteran pest Anticarsia gemmatalis (Lep.: Noctuidae) has been initially characterized following the procedures recommended by Williams and Cory. DNA hybridization and Southern blot analysis identified this isolate as a new member of the Polyiridovirus complex. The San Miguel IV MSP gene was identified and a central fragment of ca. 719 bp was recovered by PCR amplification. The restriction endonuclease profiles (5 enzymes) of this isolate were distinct from others previously described.
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PMID:Comparative studies of iridoviruses: further support for a new classification. 797 84

Acute intermittent porphyria (AIP) is an inherited disorder characterized by a deficiency of hydroxymethylbilane synthase (EC 4.3.1.8.; HMBS), the third enzyme in the heme biosynthetic pathway. To date, 113 different HMBS gene mutations have been reported in the world. However, there were a few reports of the gene mutations in the Japanese AIP patients. We studied the gene mutation in two unrelated AIP families in the San-in district, a local area of Western Japan. The overlapping 6 fragments of the HMBS gene, amplified by the reverse transcript-polymerase chain reaction, were analyzed by the single-strand conformation polymorphism with silver staining technique. The abnormal fragment from a member of one family was sequenced to detect the C to T substitution at 517 nucleic acid position of cDNA, which led to a missense mutation of arginine to tryptophan exchange at an amino acid level (R173W). This mutation located in exon 10 created a new site of the MSP 1 restriction endonuclease and was screened by the amplified fragment of exon 10 from genomic DNA with the MSP 1 digestion. The mutation was detected totally in three members of the family and interestingly also in two patients of an unrelated family. This mutation has been reported widely in the world independently, such as in a Swedish, a Canadian, a Finnish, and a French family, but is the first in Japanese patients. The screening method for this mutation is useful for diagnosis in Japanese AIP patients.
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PMID:Mutation in the exon 10 (R173W) of the hydroxymethylbilane synthase gene in two unrelated Japanese families with acute intermittent porphyria. 952 50

Although much is known about the bacterial cellulosome and its various protein components, their contributions to bacterial growth on cellulose and the process of cellulolysis in vivo cannot currently be assessed. To remedy this, the authors have developed gene transfer techniques for Clostridium cellulolyticum ATCC 35319. Firstly, transfer of Tn1545 has been obtained using an Enterococcus faecalis donor. Secondly, IncP-mediated conjugative mobilization of plasmids from Escherichia coli donors has also been achieved. The yield of transconjugants in both cases was low and was probably limited by the suboptimal growth conditions that must of necessity be employed for the co-culture of oligotrophic C. cellulolyticum with copiotrophic donors. A restriction endonuclease was detected in crude extracts of C. cellulolyticum. This enzyme, named CCE:I, is an isoschizomer of MSP:I (HPA:II). Electro-transformation was employed to establish plasmids containing the replication functions of pAMss1 (En. faecalis), pIM13 (Bacillus subtilis), pCB102 (Clostridium butyricum), pIP404 (Clostridium perfringens) and pWV01 (Lactococcus lactis subsp. cremoris) in C. cellulolyticum. Transformants were only obtained if the DNA was appropriately methylated on the external C of the sequence 5'-CCGG-3' using either BSU:FI methylase in vivo or MSP:I methylase in vitro. Plasmids based on the pAMss1 and pIM13 replicons were more stably maintained than one based on the pCB102 replicon. Selection of transformants on solid medium led to low apparent transformation efficiencies (approx. 10(2) transformants per microg DNA) which might, in part, reflect the low plating efficiency of the organism. Selection of transformants in liquid medium led to a higher apparent yield of transformants (between 10(5) and 10(7) transformants per microg DNA). The methods developed here will pave the way for functional analysis of the various cellulosome components in vivo.
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PMID:Gene transfer to Clostridium cellulolyticum ATCC 35319. 1110 65

Mapping of genomic DNA methylation is a dispensable part of functional genome. We have developed a novel method based on methylation-specific primer and serial analysis of gene expression, called MSP-SAGE, with potential of high-throughput quantification of genomic DNA methylation. We used a 6-mer methylation-specific primer to extend the methylated CpG sequences other than non-methylated CpG sequences. The 17 bp tags contained methylated CpG sequence, which were obtained from extended methylation sequence by digestion of restriction endonuclease, and then the tags were concatenated and cloned for sequencing. We can identify the locations of methylation according to the sequences of tags and quantify the methylation status from the frequency of the tags. MSP-SAGE has a good linearity in a broad methylation range from 5% to 100% with good accuracy and high precision. The proof-of-principle study shows that MSP-SAGE is a reliable high-throughput assay for quantification of DNA methylation.
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PMID:High-throughput assay of DNA methylation based on methylation-specific primer and SAGE. 1644 79

Epigenome modifications of the human genome play an important role in gene expression regulation and chromatin structure formation. Methylation of cytosine of the CpG dinucleotides is one of the main epigenone modifications of the human genome. Changes in the CpG-islands methylation status of tumor-suppressing genes and oncogenes play an important role in carcinogenesis. Determination of DNA methylation status in oncology is important for early tumor diagnostics, choice and monitoring of cancer treatment and prognosis of survival of patients with cancer after surgeries. There is a wide range of approaches to determination of DNA methylation status now. The review purpose is to characterize main approaches to the determination of the CpG dinucleotides methylation status, which could be used in oncology. They include: bisulfite sequencing, bisulfite pyrosequencing, MS-REA (methylation-sensitive restriction endonuclease assay), MSP (methylation-specific PCR), COBRA (combined bisulfite restriction analysis), MS-SnuPE (methylation-sensitive single nucleotide primer extension), MS-SSCA (methylation-sensitive single-strand conformation analysis), MethyLight, HeavyMethyl, MALDI-TOF MS (matrix-assisted laser desorption/ionization time-to-flight mass spectrometry), FMCA (fluorescence melting curve analysis), restriction genome scanning, use of microarrays for determining genes with increased expression after DNA methylation inhibition, DMH (differential methylation hybridization), use of capability of methylCpG-binding domain of MeCP2 protein to bind methylated DNA, immunoprecipitation of methylated DNA with antibodies specific for methylated cytosines. A general principle, advantages, disadvantages and potential artifacts here been described for each approach in the paper.
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PMID:[Modern methodical approaches to determining the DNA methylation status and their use in oncology]. 1914 Apr 45

Trichophyton ajelloi is a geophilic dermatophyte that specializes in the decomposition of native keratin. It exists in soil with a permanent influx of keratin matter. In the present study, two PCR-based methods were used for the identification and intra-species differentiation of T. ajelloi strains isolated from 3 types of soils with different physicochemical properties. The first method, employed for molecular identification, was PCR amplification of the 5.8S rRNA gene and its flanking regions encoding internal transcribed spacers (ITSs), followed by restriction enzyme digestion using endonuclease HinfI. The second method, employed for molecular differentiation, was microsatellite-primed PCR (MSP-PCR) using the repetitive oligonucleotide (GACA)4. All the T. ajelloi strains were also identified using a traditional culture method. Our results showed that molecular identification using the PCR-restriction fragment length polymorphism (PCR-RFLP) method agreed with the identification made using the traditional approach. On the other hand, PCR-RFLP results showed no strain differentiation, while MSP-PCR using the (GACA)4 primer identified different varieties among the T. ajelloi strains. The reasons for the intra-species differentiation of T. ajelloi have been discussed.
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PMID:Microsatellite-primed PCR for intra-species genetic relatedness in Trichophyton ajelloi strains isolated in poland from various soil samples. 2485 70