[Restriction fragment length differences of genomic repetitive DNA from five sibling species of Anopheles hyrcanus group].
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Abstract:
Genomic DNA were prepared from 5 sibling species of Anopheles hyrcanus group including An. sinensis (ASS), An. anthropophagus (ALA), An. liangshanensis (ALS), An. crawfordi (ACW) and An. xiaokuanus (AXK). High molecular weight DNA from each species were cut with three restriction endonucleases (Bgl II, Hae III and Pst I) and the DNA fragments analysed by agarose gel electrophoresis and ethidium bromide staining. Three enzymes (Bgl II, Hae III and Pst I) produced unique fragments in all sibling species tested. A diagnostic restriction fragment length of DNA from 5 sibling species of An. hyrcanus group may be derived from a single restriction enzyme pattern (i.e. Bgl II). The results demonstrated that the restriction fragment length differences of repetitive DNA could used as a tool to distinguish sibling species of An. hyrcanus group.Keywords:
Restriction fragment
genomic DNA
Ethidium bromide
Agarose gel electrophoresis
Restriction site
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Substance P (SP) is an important neuropepetide detected in a variety of locations in the central nervous system. Variations in SP content or SP receptors in psychiatric disorders have been described. Using SP clones as probes the authors have found three restriction fragment length polymorphisms (RFLPs) in the SP gene. The RFLPs are generated by digestion of genomic DNA with the MspI, and RsaI and NcoI restriction endonucleases. The MspI RFLP is detected by two genomic clones mapping to the 5' end of the gene while the RsaI and NcoI rFLPs are both detected by two genomic clones on the 3' end and also by a full-length cDNA clone of the gene. All three RFLPs are characterized by two alleles. For the MspI RFLP the frequency of both alleles is similar, for the Rsa I and NcoI RFLP one of the alleles is significantly more abundant than the other. These RFLPs are now being used to determine whether any of the alleles correlate with either schizophrenia or affective disorder.
genomic DNA
Restriction fragment
Restriction map
clone (Java method)
Restriction site
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A genomic library of partially EcoRI-digested DNA from the lesser snow goose, Anser caerulescens caerulescens, was constructed in the phage vector Charon 4. Phage containing only unique sequences were identified by screening plaques with 32P-labeled genomic DNA. Restriction-fragment-length polymorphisms (RFLPs) were identified by probing DNA from 11-13 male birds from the breeding colony at La Perouse Bay. Of the 17 probes examined, all detected RFLPs with at least one of EcoRi, HindIII, Msp1, and Taq1. Several of them identified highly variable regions with multiple alleles. These RFLPs are valuable DNA markers that can be used for (1) the examination of DNA variation, relatedness, and genetic distance and (2) assessing paternity and maternity. These data suggest that there are higher levels of variation of DNA sequence in birds than had previously been thought to exist.
Minisatellite
genomic DNA
Restriction fragment
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We studied the polymorphism of HLA‐DR2 by Southern blot analysis. Genomic DNA from 12 DR2 positive unrelated individuals was digested separately with five restriction endonucleases, Bam HI, Eco RI, Eco RV, Hind III and Pvu II. Hybridizing restriction fragments were visualized using radiolabeled β‐chain cDNA probes homologous to the DR and DQ subregions of the HLA. The results in the present study demonstrate that the two subtypes of DR2, DR2.1 and DR2.2, as defined by serological and cellular (PLT) techniques can be identified by Southern blot analysis. These two subtypes were identifiable after cleaving genomic DNA with Bam HI and EcoRV. DR2.1 correlated with restriction endonuclease fragments at 2.8 kb (DRβ) and 6.6 kb (DQβ) after Bam HI digest, while DR2.2 correlated with a 2.9 kb (DRβ) and a 2.6 kb (DQβ) restriction fragment after Eco RV digest. In addition, inheritance of the restriction fragment lengths defining DR2.1 and DR2.2 were observed in two families. In contrast, no differences between DR2.1 and DR2.2 were observed with the restriction enzymes Eco RI, Hind III and Pvu II.
genomic DNA
Southern blot
Restriction fragment
Restriction map
EcoRV
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The present study was initiated to focus on the differences or similarities among different isolates of Trypanosoma evansi through restriction endonuclease profile. The genomic DNA of T.evansi, isolated from naturally infected buffalo, horse and camel were analysed. A panel of restriction enzymes-Alu I, Dra I, EcoR I, Hind III, Kpn I, Not I, Pst I, Sal I, Sma I and Taq I were used for complete digestion of genomic DNA. The agarose gel electrophoresis of digested DNA samples appeared as continuous smear along the electrophoretic tracks on ethidium bromide staining revealing the complex size oftrypanosome genome. There was no fixed restriction site, but in restriction enzyme Dra I and Alu I where restriction site at the region of 1.5 kb and 100 bp, respectively, appeared with background smear of DNA fragments. No heterogeneity in the nuclear DNA restriction endonuclease profile among the isolates was recorded.
Trypanosoma evansi
Ethidium bromide
genomic DNA
Agarose gel electrophoresis
Restriction fragment
Restriction map
Restriction site
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Five restriction endonucleases were used to digest genomic DNA from 5 isolates of Trichinella spiralis obtained from Changchun, Tianjin, Xian, Henan and Yunnan. All the isolates were secured from pigs except the Changchun strain which came from dog. The DNA fragments digested by endonuclease were separated by agarose gel electrophoresis. The Changchun isolate had a EcoRI band at 1.12kb and a DraI band at 1.97kb which were unique to this isolate. A cloned specific repetitive DNA sequence (1.12kb) from the Changchun strain was selected to prepare a probe for the Southern blotting of EcoRI restriction DNA fragments for the 5 isolates. The 1.12kb hybridizing band did not appear except in the Changchun isolate. These results seem to indicate that there are differences between the isolates obtained from hosts in different geographical regions.
Trichinella spiralis
genomic DNA
Southern blot
Agarose gel electrophoresis
Restriction fragment
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Five restriction endonucleases were used to digest genomic DNA from 5 isolates of Trichinella spiralis obtained from Changchun,Tianjin,Xian,Henan and Yunnan.All the isolates were secured from pigs ex-cept the Changchun strain which came from dog.The DNA fragments digested by endonuclease were sepa-mted by agarose gel electrophoesis.The DNA fragments digested by endonuclease were sepa-rated by agarose gel electrophoresis.The Changchun is olate had a EcoRI band at 1.12kb and a Dral band at 1.97kb which were unique to this isolate.A cloned specific repetitive DNA sequence(1.12kb) from the Changchun strain was selected to prepare a probe for the Southern blotting of EcoRI restriction DNA frag-ments for the 5 isolates.The 1.12kb hybridizing band did not appear except in the Changchun isolate.These results seem to indicate that there are differences between the isolates obtained from hosts in differ-ent geographical regions.
Trichinella spiralis
genomic DNA
Agarose gel electrophoresis
Southern blot
Restriction fragment
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Genomic DNA were prepared from 5 sibling species of Anopheles hyrcanus group including An. sinensis (ASS), An. anthropophagus (ALA), An. liangshanensis (ALS), An. crawfordi (ACW) and An. xiaokuanus (AXK). High molecular weight DNA from each species were cut with three restriction endonucleases (Bgl II, Hae III and Pst I) and the DNA fragments analysed by agarose gel electrophoresis and ethidium bromide staining. Three enzymes (Bgl II, Hae III and Pst I) produced unique fragments in all sibling species tested. A diagnostic restriction fragment length of DNA from 5 sibling species of An. hyrcanus group may be derived from a single restriction enzyme pattern (i.e. Bgl II). The results demonstrated that the restriction fragment length differences of repetitive DNA could used as a tool to distinguish sibling species of An. hyrcanus group.
Restriction fragment
genomic DNA
Ethidium bromide
Agarose gel electrophoresis
Restriction site
Cite
Citations (1)
Restriction fragment length polymorphisms in the ribosomal DNA (rDNA) have been shown to be a useful criterion for distinguishing among various isolates of Candida albicans. In a sample of 12 clinical isolates, we found six different classes based on variations in the fragments produced from genomic DNA by EcoRI and visualized after Southern transfer by being probed with a plasmid containing Saccharomyces cerevisiae rDNA. Some of the classes appeared to be heterozygous at the rDNA locus. Similar digestion of other Candida species showed that each could be identified on the basis of its restriction patterns. Since these are highly reiterated genes, the differences were apparent on ethidium bromide-stained gels; Southern transfers were not necessary. EcoRI restriction maps of the rDNA of C. albicans, C. stellatoidea, C. tropicalis, and C. guilliermondii were determined.
Ethidium bromide
Restriction fragment
genomic DNA
Southern blot
Restriction site
Restriction map
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Using labelled, γ-32P rRNA of mycobacteria as a probe restriction fragment length polymorphism (RFLP) of rRNA genes of strains belonging to the Mycobacterium fortuitum-chelonei complex was analysed. Each DNA sample was cleaved with EcoRI restriction endonuclease, the fragments were separated by agarose gel electrophoresis and transferred to nitrocellulose membrane. Fragments of DNA containing rRNA genes were identified by hybridization with γ-32P-labelled rRNA. Patterns were found to be species specific and both the species were distinguishable from each other. Results indicate that this approach can be used for rapid genomic characterization of the Mycobacterium fortuitum-chelonei complex.
Mycobacterium fortuitum
Agarose gel electrophoresis
genomic DNA
Restriction fragment
5S ribosomal RNA
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Citations (8)
PstI
HaeIII
genomic DNA
EcoRV
Restriction site
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Citations (248)