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)

Genomic DNA from Schistosoma mansoni and S. japonicum adult worms was hybridized to a 32p-labelled pSM 889 probe after cleavage by restriction endonuclease BglII, BamHI, XbaI and EcoRI, or to a 32p-labelled pSM 389 probe after cleavage by EcoRI. The resulting hybridization banding patterns were significantly different between the two species. Genomic DNA from S. japonicum adult worms of Hunan, Hubei, Jiangxi, Zhejiang and Yunnan isolates was hybridized to a 32p-labelled pSM 389 probe after cleavage by EcoRI. The major hybridization bands were identical while the minor bands were more or less different among the isolates. Of them, isolates Hunan, Hubei and Zhejiang have similar minor bands while isolates Jiangxi and Yunnan have minor bands significantly different from those of the above three isolates and also markedly distinct from those of each other. These results indicate that the major hybridization banding patterns are unique to schistosome species while the minor banding patterns may serve as the basis for strain differentiation.
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PMID:[Differentiation of schistosome species and strains by DNA hybridization]. 840 68

We describe a variety of restriction site polymorphisms in the introns of Ascaris nuclear genes and in the ribosomal DNA spacers. We use these markers, in addition to previously described mitochondrial variation, to clarify our understanding of the epidemiology of Ascaris in Guatemalan villages where humans and pigs occur in sympatry and to describe the genetic structure of host-associated Ascaris populations from world-wide locations. Intron sequences were amplified from individual worms and alleles defined by endonuclease digestion. Two loci were monomorphic, while 4 length variants and 22 point mutations were detected in the other 7 loci. Within sympatric Guatemalan populations no single locus from either the nuclear or mitochondrial genome was fixed for alternative alleles, although allele frequencies were significantly different at many loci. Phenograms constructed from multilocus nuclear genotypes of individual worms failed to reveal a single case of cross-infection, and demonstrate that divergent mtDNA genotypes are segregating within host-associated populations. On a world-wide scale, the data suggest that extant worm populations result from a single host shift, although characterization of genetic variation in additional loci will be necessary to confirm this. The direction and the geographical origin of the host shift were unresolved. Overall 65% of nuclear genetic variation was found within populations, host (human or pig) explained 18%, while geographical variation within host-associated populations explained 17%. The results (a) demonstrate the utility of introns for studying the epidemiology of parasites showing limited allozyme variation (b) suggest that programmes aiming to control Ascaris infection in the human population can safely ignore zoonotic infection from pigs and (c) illustrate the problems inherent in using single genetic markers to make inferences about the epidemiology of closely related parasite taxa.
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PMID:Host specificity, evolutionary relationships and macrogeographic differentiation among Ascaris populations from humans and pigs. 930 Apr 71

The polymerase chain reaction was used to amplify fragments comprising the known reading frame of the nematode nicotinic acetylcholine alpha-subunit gene tar-1. Sequences were derived from DNA prepared from bulk collections of worms and from individual male and female Trichostrongylus colubriformis. In each case a levamisole-resistant (BCk) and a drug susceptible population were examined. Although several nucleotide transitions were detected no amino acid sequence variations were found between the isolates and between individual worms, indicating that the coding sequence of this gene is not responsible for levamisole-resistance in the isolate tested. However, an intronic allelic T/C variation at position 4955 was observed in both populations. It has been reported that levamisole-resistance in the BCk isolate of T. colubriformis is due to a sex-linked recessive gene or gene complex. A restriction fragment length polymorphism formed by the allelic variation was found and was detectable by digestion with the restriction endonuclease NlaIII. Statistical comparison of allele frequencies from individual male and female worms was consistent with sex-linkage of tar-1 (P < 0.05) but showed no correlation with levamisole resistance status. The polymorphism described will provide a useful X-chromosome marker and represents the first mapped genetic locus in this species.
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PMID:The nicotinic acetylcholine alpha-subunit gene tar-1 is located on the X chromosome but its coding sequence is not involved in levamisole resistance in an isolate of Trichostrongylus colubriformis. 947 89

In this study, we assessed single-strand conformation polymorphism (SSCP)-based approaches for their capacity to fingerprint sequence variation in ribosomal DNA (rDNA) of ascaridoid nematodes of veterinary and/or human health significance. The second internal transcribed spacer region (ITS-2) of rDNA was utilised as the target region because it is known to provide species-specific markers for this group of parasites. ITS-2 was amplified by PCR from genomic DNA derived from individual parasites and subjected to analysis. Direct SSCP analysis of amplicons from seven taxa (Toxocara vitulorum, Toxocara cati, Toxocara canis, Toxascaris leonina, Baylisascaris procyonis, Ascaris suum and Parascaris equorum) showed that the single-strand (ss) ITS-2 patterns produced allowed their unequivocal identification to species. While no variation in SSCP patterns was detected in the ITS-2 within four species for which multiple samples were available, the method allowed the direct display of four distinct sequence types of ITS-2 among individual worms of T. cati. Comparison of SSCP/sequencing with the methods of dideoxy fingerprinting (ddF) and restriction endonuclease fingerprinting (REF) revealed that also ddF allowed the definition of the four sequence types, whereas REF displayed three of four. The findings indicate the usefulness of the SSCP-based approaches for the identification of ascaridoid nematodes to species, the direct display of sequence variation in rDNA and the detection of population variation. The ability to fingerprint microheterogeneity in ITS-2 rDNA using such approaches also has implications for studying fundamental aspects relating to mutational change in rDNA.
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PMID:Single-strand conformation polymorphism (SSCP)-based mutation scanning approaches to fingerprint sequence variation in ribosomal DNA of ascaridoid nematodes. 969 83

A mitochondrial NADH dehydrogenase I gene fragment (NDI) was sequenced for three laboratory maintained isolates of Schistosoma japonicum. Comparison of sequences representing the isolates (originally obtained from the Anhui and Zhejiang provinces of the People's Republic of China, and from the Philippines) revealed inter-isolate sequence variations of 0.2-0.6% and no intra-isolate variation was found. The sequences also indicated that while the amplification products of the Zhejiang and Philippine isolates contained a recognition site for the endonuclease RsaI, there was no such site in the Anhui isolate. This was tested by digesting amplification products from a number of individual worms with RsaI. Then an infection experiment was designed to test the value of this genetic marker for studies of the population biology of S. japonicum in the final host. For this, the two Chinese isolates were used. Three groups of mice (A-C) were exposed firstly to a primary infection and then challenge-infected at weeks 4 and 7 of the experiment. In group A the first infection was done with the Anhui isolate, and the two others with the Zhejiang isolate, thereby providing a specific, detectable cohort. In groups B and C the Anhui isolate was used for the second and third infection. All mice were perfused 5 weeks after the last challenge infection, and the NDI was subsequently amplified from DNA of the perfused worms and digested with RsaI. The digestion revealed that while infection groups A and B contained mixed populations of the Anhui and Zhejiang isolates, only Zhejiang worms were present in group C. We concluded that the absence/presence of the RsaI site in the NDI provides a useful marker for the delineation of cohorts of S. japonicum.
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PMID:PCR-based identification of individuals of Schistosoma japonicum representing different subpopulations using a genetic marker in mitochondrial DNA. 1050 22

The Caenorhabditis elegans nuc-1 gene has previously been implicated in programmed cell death due to the presence of persistent undegraded apoptotic DNA in nuc-1 mutant animals. In this report, we describe the cloning and characterization of nuc-1, which encodes an acidic nuclease with significant sequence similarity to mammalian DNase II. Database searches performed with human DNase II protein sequence revealed a significant similarity with the predicted C. elegans C07B5.5 ORF. Subsequent analysis of crude C. elegans protein extracts revealed that wild-type animals contained a potent endonuclease activity with a cleavage preference similar to DNase II, while nuc-1 mutant worms demonstrated a marked reduction in this nuclease activity. Sequence analysis of C07B5.5 DNA and mRNA also revealed that nuc-1(e1392), but not wild-type animals contained a nonsense mutation within the CO7B5.5 coding region. Furthermore, nuc-1 transgenic lines carrying the wild-type C07B5.5 locus demonstrated a complete complementation of the nuc-1 mutant phenotype. Our results therefore provide compelling evidence that the C07B5.5 gene encodes the NUC-1 apoptotic nuclease and that this nuclease is related in sequence and activity to DNase II.
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PMID:The C. elegans apoptotic nuclease NUC-1 is related in sequence and activity to mammalian DNase II. 1090 46

Schistosoma mansoni genomic DNA from male and female adult worms was subjected to restriction by the isoschizomeric endonucleases HpaII and MspI, which display different sensitivities with respect to cytosine methylation. The digested DNA was hybridized with 13 S. mansoni probes. Southern blot analysis showed that there were no observable differences in the restriction patterns of the two isoschizomers and that the patterns were identical in male and female parasites. Adenine methylation was also ruled out since no differences were observed with DpnI, Sau3A1, or MboI restriction enzymes. The methylation-dependent restriction endonuclease McrBC, which cleaves DNA containing methylcytosine and will not cleave unmethylated DNA, did not digest S. mansoni genomic DNA. These results demonstrate that the genome of adult S. mansoni is not methylated.
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PMID:Lack of DNA methylation in Schistosoma mansoni. 1152 39

A 474 bp fragment of the mitochondrial cytochrome oxidase c subunit 1 (cox1) of Cooperia oncophora was cloned and sequenced. The overall nucleotide diversity of the cox1 fragment varied from 0.5 to 2.0% between individuals. Two nucleotide substitutions were found within two RsaI endonuclease restriction sites and were used in a PCR-based restriction fragment length polymorphism (PCR-RFLP) assay to asses the intra-population variation of C. oncophora. Testing 816 individuals revealed the existence of three different haplotypes, having either both (type I) or only one (types II and III) RsaI site. Laboratory maintained individuals obtained at different time points after infection showed no significant difference in the distribution of the three haplotypes. Neither was there a difference in the distribution between male and female worms, confirming that the mitochondrial genome of C. oncophora is also maternally inherited. Nevertheless, there was a significant difference in the prevalence of the RsaI point mutation in the cox1 gene between the laboratory maintained population of C. oncophora and a Dutch field isolate, indicating that these RFLPs can be used to study genetic variation within or among C. oncophora populations.
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PMID:Cytochrome oxidase c subunit 1 polymorphisms show significant differences in distribution between a laboratory maintained population and a field isolate of Cooperia oncophora. 1455 66

In the present study, samples of Oesophagostomum spp. collected from pigs from different geographical localities in mainland China were characterized genetically by polymerase chain reaction-linked single-strand conformation polymorphism (PCR-SSCP) and restriction fragment length polymorphism (PCR-RFLP) techniques using genetic markers in the internal transcribed spacers (ITS) of nuclear ribosomal DNA (rDNA). The second internal transcribed spacer (ITS-2) was amplified from 51 individual nodule worms by PCR, and the amplicons were analyzed by SSCP. With the exception of slight microheterogeneity, SSCP analyses displayed two distinct banding profiles that allowed the identification of all Oesophagostomum spp. samples examined into two groups, the first one represented O. dentatum, and the second one may represent O. quadrispinulatum. Then, the entire ITS was amplified from individual samples, and the amplicons were digested with restriction endonuclease Pst I. The results of RFLP analyses were consistent with that of SSCP. Sequence analysis of ITS rDNA supported the identification and differentiation of Chinese Oesophagostomum spp. samples into two species, namely, O. dentatum and O. quadrispinulatum. These PCR-based approaches provide useful complementary tools to traditional methods for the accurate identification of Oesophagostomum spp. (irrespective of developmental stage) and have implications for studying the ecology and population genetic structures of these parasites and for the prevention and control of the diseases they cause.
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PMID:Characterization of Oesophagostomum spp. from pigs in China by PCR-based approaches using genetic markers in the internal transcribed spacers of ribosomal DNA. 1731 77

We previously showed that Caenorhabditis elegans APN-1, the only metazoan apurinic/apyrimidinc (AP) endonuclease belonging to the endonuclease IV family, can functionally rescue the DNA repair defects of Saccharomyces cerevisiae mutants completely lacking AP endonuclease/3'-diesterase activities. While this complementation study provided the first evidence that APN-1 possesses the ability to act on DNA lesions that are processed by AP endonucleases/3'-diesterase activities, no former studies were conducted to examine its biological importance in vivo. Herein, we show that C. elegans knockdown for apn-1 by RNAi displayed phenotypes that are directly linked with a defect in maintaining the integrity of the genome. apn-1(RNAi) animals exhibited a 5-fold increase in the frequency of mutations at a gfp-lacZ reporter and showed sensitivities to DNA damaging agents such as methyl methane sulfonate and hydrogen peroxide that produce AP site lesions and strand breaks with blocked 3'-ends. The apn-1(RNAi) worms also displayed a delay in the division of the P1 blastomere, a defect that is consistent with the accumulation of unrepaired lesions. Longevity was only compromised, if the apn-1(RNAi) animals were challenged with the DNA damaging agents. We showed that apn-1(RNAi) knockdown suppressed formation of apoptotic corpses in the germline caused by an overburden of AP sites generated from uracil DNA glycosylase mediated removal of misincorporated uracil. Finally, we showed that depletion of APN-1 by RNAi partially rescued the lethality resulting from uracil misincorporation, suggesting that APN-1 is an important AP endonuclease for repair of misincorporated uracil.
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PMID:Caenorhabditis elegans APN-1 plays a vital role in maintaining genome stability. 2003


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