Gene/Protein Disease Symptom Drug Enzyme Compound
Pivot Concepts: Gene/Protein Disease Symptom Drug Enzyme Compound   Target Concepts: 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)

Among a mixture of amphotropic and ecotropic murine leukemia viruses (MuLVs) isolated from paralyzed wild mice, only N-tropic ecotropic MuLV, cloned by cell culture techniques, has been shown to induce paralysis after reinjection into susceptible mice (M. B. Gardner, Curr. Top. Microbiol. Immunol. 79:215-239, 1978). The viral DNA genome of one of these neurotropic MuLVs (Cas-Br-E) has been cloned in Charon 21A at the SalI site. One clone, designated NE-8, was studied in more detail. A restriction endonuclease map of this cloned DNA was derived. Cloned viral DNA microinjected into NIH 3T3 cells produced infectious MuLV which was characterized as XC+, ecotropic, and N-tropic. The virus that was recovered after the microinjection of NE-8 DNA was also injected into susceptible SIM.S and NIH Swiss mice and was found to induce lower limb paralysis in these animals. These results make it highly unlikely that other agents (which might have escaped detection and separation from ecotropic MuLV by the techniques previously used) play a role in the etiology of this disease and clearly indicate that the ecotropic MuLV genome harbors sequences responsible for this paralysis. The availability of this clone DNA would now allow us to map these sequences on the genome.
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PMID:Molecular cloning of infectious viral DNA from ecotropic neurotropic wild mouse retrovirus. 630 Apr 52

Two new isolates of cricket paralysis virus, TAR and SIM, are described that were originally isolated from laboratory colonies of Drosophila melanogaster and Drosophila simulans respectively. Using a combination of biological, serological and molecular characters it was possible to distinguish the SIM isolate from all other isolates and it is thus described as a new strain; CrPVSIM. The TAR isolate however, was indistinguishable from the CrPV reference isolate CrPVVIC/GM/D2(2)/Gm/D2(2) (Teleogryllus commodus, Victoria, Australia, 1968). The molecular characters used in the present study were obtained by combining PCR and restriction endonuclease digestion of the amplified fragments. This work demonstrates that such molecular characters, when used in combination with others, provide a powerful set of taxonomic characters for classifying CrPV isolates and strains and assessing their genetic relatedness.
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PMID:A molecular taxonomy for cricket paralysis virus including two new isolates from Australian populations of Drosophila (Diptera: Drosophilidae). 885 30

Labeling and tracing of specific sequences in living cells has been a major challenge in studying the spatiotemporal dynamics of native chromatin. Here we repurposed the prokaryotic CRISPR/Cas adaptive immunity system to specifically detect endogenous genomic loci in mouse embryonic stem cells. We constructed a catalytically inactive version of the Cas9 endonuclease, fused it with eGFP (dCas9-eGFP) and co-expressed small guide RNAs (gRNAs) to target pericentric, centric, and telomeric repeats, which are enriched in distinct nuclear structures. With major satellite specific gRNAs we obtained a characteristic chromocenter (CC) pattern, while gRNAs targeting minor satellites and telomeres highlighted smaller foci coinciding with centromere protein B (CENP-B) and telomeric repeat-binding factor 2 (TRF2), respectively. DNA sequence specific labeling by gRNA/dCas9-eGFP complexes was directly shown with 3D-fluorescent in situ hybridization (3D-FISH). Structured illumination microscopy (3D-SIM) of gRNA/dCas9-eGFP expressing cells revealed chromatin ultrastructures and demonstrated the potential of this approach for chromatin conformation studies by super resolution microscopy. This programmable dCas9 labeling system opens new perspectives to study functional nuclear architecture.
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PMID:Visualization of specific DNA sequences in living mouse embryonic stem cells with a programmable fluorescent CRISPR/Cas system. 2463 35

Identifying cis-regulatory elements is critical in understanding the direct and indirect interactions that occur within gene regulatory networks. Current approaches include DNase-seq, a technique that combines sensitivity to the nonspecific endonuclease DNase I with high-throughput sequencing to identify regions of regulatory DNA on a genome-wide scale. Yet, challenges still remain in processing recalcitrant tissues that have low DNA content. Here, we describe DNase I SIM (for Simplified In-nucleus Method), a protocol that simplifies and facilitates generation of DNase-seq libraries from plant tissues for high-resolution mapping of DNase I hypersensitive sites. By removing steps requiring the use of gel agarose plugs in DNase-seq, DNase I SIM reduces the time required to perform the protocol by at least 2 days, while also making possible the processing of difficult plant tissues including plant roots.
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PMID:DNase I SIM: A Simplified In-Nucleus Method for DNase I Hypersensitive Site Sequencing. 2862 84

The worldwide spread of carbapenemase producing Enterobacteriaceae isolates has become a major threat of public health. This worrisome situation leads the development of new methods for carbapenemase screening, detection, prevention of spread and epidemiological data collection as mandatory. In this study, it was aimed to investigate existence and distribution of carbapenemase-encoding genes (CEGs) among carbapenem-resistant Enterobacteriaceae isolated from various clinical samples in Ankara University Faculty of Medicine, Ibni Sina Hospital, Central Microbiology Laboratory between June 2010-May 2014 and detect their clonal relationship. A total of 112 non-repetitive Enterobacteriaceae isolates which were intermediate or resistant to ertapenem were identified by using Phoenix (BD Diagnostic Systems, Sparks, USA) automated microbiology system. After DNA extraction from the isolates, 11 carbapenemase-encoding genes (CEGs) (bla_IMP, bla_VIM, bla_SPM, bla_KPC, bla_NDM, bla_OXA-48, bla_GIM, bla_SIM, bla_AIM, bla_DIM ve bla_BIC) were detected with PCR. The clonal relationship among the isolates was determined by PFGE method following digestion with Xbal DNA macrorestriction endonuclease. Among 112 isolates Klebsiella pneumoniae was the most frequent (n= 79, 70.5%) bacteria followed by Escherichia coli (n= 15, 13.4%), Enterobacter cloacae (n= 10, 8.9%), Enterobacter aerogenes (n= 4, 3.6%) and Klebsiella oxytoca (n= 4, 3.6%) respectively. bla_OXA-48 was the most frequent gene detected. Among 83 (74.1%) isolates bla_OXA-48 was detected alone and in 7 (6.3%) of the isolates it was identified with bla_VIM gene coexistence. bla_VIM gene was identified as the second most frequent CEG among the isolates. bla_VIM gene was detected positive in 9 (8%) isolates. bla_NDM gene was identified in 2 (1.8%) isolates. Ten of the K.pneumoniae isolates with identical PFGE pattern were named as pulsotype B. These isolates were found to be similar in terms of isolate location, isolation dates, antibiotic resistance patterns and the carbapenemase genes they carry, and are considered to be potential outbreak isolates originated from intensive care units. On the other hand CEGs were found in the clinical samples obtained from five out-patients suggesting that community-acquired infections may also arise due to carbapenemase producing Enterobacteriaceae in our country where blaOXA-48 producers are endemic. According to this study, bla_OXA-48 producing gram negative bacteria were frequent in our hospital. The prevalance of bla_VIM gene among metallo-beta-lactamases and coexistence with bla_OXA-48 gene was remarkable. The frequency of blaNDM producing isolates in our hospital was not detected as high yet. In this study, the identification of carbapenemase producing bacteria as outbreak strains in our hospital indicated that cross-sectional surveillance for carbapenemase-producing bacteria from each patient was valuable in terms of early diagnosis of outbreaks.
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PMID:[Investigation of carbapenemase genes and molecular epidemiology of Enterobacteriaceae strains isolated between 2010-2014 in a university hospitals]. 2964 25

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