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)

Nick translation labelling technique was applied to the preparation of two 32P labelled genomic DNA probes from L. interrogans serovar Lai strain 017 and Leptonema illini strain 3055, respectively. Dot-blotting and Southern-blotting with DNA of five strains leptospira from different genus and species were performed. The results showed that certain differences among the L. interrogans, L. biflexa and Leptonema illini could be detected by endonuclease assay. L. interrogans strain 017 with L. biflexa strain Patoc I and Leptonema illini 3055 strain exhibited very little homology. L. interrogans strains 017, 601, and 245 from different serogroup and serovar had a high degree of homology. Leptonema illini strain 3055 showed little homology with L. interrogans and L. biflexa. Therefore DNA hybridization may be used as a tool for the identification and classification of leptospira.
Hua Xi Yi Ke Da Xue Xue Bao 1992 Jun
PMID:[Assay of genomic DNA homology among strains of different virulent leptospira by DNA hybridization]. 145 38

For the application of restriction endonuclease analysis in typing and identifying leptospira, we selected some serovars and isolates, and analysed preliminarily their DNA with four restriction enzymes, EcoR I, Bgl II, Hha I, and Hind III. The DNA samples were isolated from the reference strains and isolates as follows: Serovar lai 56601, 017 (the virulent strain for PDH model of guinea pig), Serovar autumnalis 56606, Serovar manhao II 67020, and isolates 87112 and 87369. Each 2 micrograms of DNA was digested with 20mu of restriction enzyme at 37 degrees C for 2h and electrophoresed in 0.8% agarose gel. The gels were stained in ethidium bromide and photographed with UV light. In our experiments, apparently different restriction patterns of serovar lai 017 were observed with four restriction enzymes. Serovar lai, serovar autumnalis and serovar manhao II showed different patterns with EcoR I, especially in high molecular regions. We also observed in serovar lai 017 a distinct 10.5kb band which was obscure in 56601, the reference strain of serovar lai, after EcoR I digestion. The three serovars showed some delicate differences in Hind III restriction pattern. The two isolates from Apodemus agrarius in Sichuan (1987) 87112 and 87369 had patterns identical to those of serovar lai 56601, 017 with EcoR I, and 87112 also had a pattern identical to 56601, 017 with Hind III. Our results indicate that selected three serovars can be identified by analysis of their DNA with EcoR I and Hind III. It is suggested that restriction endonuclease analysis be a good method in typing and identify leptospira and in studying the differences of special DNA molecules.
Hua Xi Yi Ke Da Xue Xue Bao 1991 Sep
PMID:[Analysis of leptospiral deoxyribonucleic acids by restriction endonucleases]. 166 Aug 46

The aim of the present study was to investigate the potential of restriction endonuclease analysis (REA) for the identifying and typing of Actinobacillus actinomycetemcomitans (Aa). Bacterial chromosomal DNA was extracted and digested with BamHI, HindIII, Sal I and Xho I respectively and separated by agarose gel electrophoresis. DNA fragment patterns of Aa and Haemophilus aphrophilus differed strikingly from each ether. The patterns of Aa FDC Y4 and Aa ATCC 29523, belonging to two different serotypes, also differed notably. Among the 4 enzymes used, Sal I and Xho I gave the most satisfactory results. The results of our work suggested that REA could be used to identify and type Aa. REA is stable, sensitive, accurate and reproducible. The method might be valuable in molecular epidemiologic studies on periodontopathic organisms.
Hua Xi Yi Ke Da Xue Xue Bao 1990 Sep
PMID:[Identification and typing of Actinobacillus actinomycetemcomitans by restriction endonuclease analysis]. 196 93

Four of positive clones were obtained by three runs of screening the cDNA library of human liver with rabbit antihuman apolipoprotein (apo) CIII IgG. One of them was identified to be the positive clone of apoC III cDNA by restriction endonuclease map studies of the cDNA insert fragment and by Western blot analysis of the expression products. The apoCIII cDNA-gamma gt11 recombinant DNA prepared from the positive clone can be used in RNA-dot blot analysis as apoCIII cDNA probe. And the apoCIII cDNA clone can be also used to produced apoCIII peptide under the induction of isopropyl-beta-D-thiogalactoside. The apoCIII peptide, after purification, has a lot of use in further studies.
Hua Xi Yi Ke Da Xue Xue Bao 1990 Sep
PMID:[Screening and identification of apolipoprotein CIII cDNA clones from human liver cDNA library and preparation of apolipoprotein CIII cDNA probe from the clones]. 209 28

Determination of genome size is very important in genetic studies and the molecular cloning of leptospires. With pulsed field gel electrophoresis (PFGE), which can be used in the analysis of large DNA molecules, we studied the genome size of leptospires and analysed it with rare cutter restrict endonuclease Not I. The PFGE system used was CHEF-DR II which produced parallel bands in agarose gel. The results showed that the genome size of leptospires was 2000kb and there was no dramatic difference between the 5 strains of 2 genus leptospira. Rare cutter Not I digestion of genome DNA of leptospira resulted in 11 bands and its bands pattern was quite different from that of E. coli, S. choleraesuis, S. typhimurium, and S. dysenteriae genome DNA. It is thought that more information can be obtained about leptospiral DNA with PFGE technique.
Hua Xi Yi Ke Da Xue Xue Bao 1990 Sep
PMID:[The study on genome size of leptospires]. 209 31

The DNAs of different serogroups and serovars of leptospires were analyzed with seven kinds of restriction endonuclease (Bam HI, Bgl II, Cfo I, Eco RI, Hind III, Kpn I and Pst I). The results showed that there are significant differences among the leptospiral DNAs of different serovars, especially in the high molecular-weight region, but the patterns of different strains belonging to the same serovar are similar. In addition, Cfo I and Eco RI are more effective for differentiating leptospires.
Hua Xi Yi Ke Da Xue Xue Bao 1993 Sep
PMID:[Restriction endonuclease analysis of leptospiral DNA from different serogroup and serovar]. 828 92

This is a study aimed at the molecular mechanisms of beta-defensins gene expression and it's gene transfer experiments for preventing mucosal infection. A rat rBD-1 cDNA was cloned. The total RNA was isolated from rat kidney. The cDNA fragment was amplified by RT-PCR with specific primers. The purified RT-PCR product was cloned in pGEM-T Easy vector. The results of restriction endonuclease pattern analysis of the recombinant plasmid, DNA sequencing, and aligning of the putative amino acid sequence with hBD-1 and mBD-1 demonstrated that rat beta-defensin rBD-1 gene was cloned successfully.
Hua Xi Yi Ke Da Xue Xue Bao 1999 Dec
PMID:[Cloning of rat beta-defensin rBD-1 cDNA]. 1138 38

New fluorogenic probes for restriction endonucleases based on molecular beacons were developed. These hairpin probes were single-stranded oligonucleotides that had stem-and-loop structure and carried 5'- fluorophor moiety and 3'-quencher moiety. Their stem sequences were designed as recognition sites for restriction endonucleases and loop sequences were unrelated nucleotides. Upon cleavage by endonuclease, these probes became fluorescent and thus could trace the enzyme activity continuously. Two probes were designed for BglII and NcoI, respectively, and each was labeled with a fluorophor of different color. The results showed that the two probes could specifically assay for the corresponding enzymes either individually or simultaneously in a real-time mode. Considering the simplicity, quantification and high throughput, these probes could be extended to other applications such as drug screening, protein-nucleic acid interaction study and searching for small molecule DNA cleavage agents.
Sheng Wu Hua Xue Yu Sheng Wu Wu Li Xue Bao (Shanghai) 2002 May
PMID:[Hairpin probes for teal-time assay of restriction endonucleases]. 1201 46

The human DNA fragmentation factor (DFF) is a heterodimer of 40 kD and 45 kD subunits. The 40 kD subunit (DFF40) has an intrinsic DNase activity responsible for the genomic DNA degradation into nucleosomal fragments during apoptosis. As an inhibitor for DFF40, the 45 kD subunit (DFF45) complexes with DFF40, inhibiting DNase activity until certain apoptosis signals are received. In cells undergoing apoptosis, the cleavage of DFF45 by activated caspase-3 frees DFF40from the complex and initiates the apoptosis-specific DNA fragmentation. In this report, the coding region of human DFF45 gene was amplified from the total RNA of HeLa cells by RT-PCR. The resulting 1 kb DNA fragment was cloned into the bacterial expression vector pET-28a(+) with a 6xhistidine tag fused to the N-terminus of DFF45, generating plasmid pET28a-DFF45, which was then used to transform E.coli BL21(DE3). Induced by IPTG, the recombinant DFF45 was expressed efficiently with a yield of 56.6% of total bacterial proteins. The product was purified to homogeneity through a nickel affinity column, followed by heat treatment, and approximately 4--6 mg of DFF was purified from 100 ml culture. Purified recombinant human DFF45, added into the apoptotic cell-free system of Xenopus egg extracts, could effectively inhibit both the digestion of lambdaDNA and the degradation of chromosomal DNA into nucleosomal fragments in the nuclei of chicken red blood cells. Our results demonstrated the existence of an apoptosis-specific endonuclease in this cell-free system, the activity of which could be inhibited by recombinant human DFF45.
Sheng Wu Hua Xue Yu Sheng Wu Wu Li Xue Bao (Shanghai) 2001
PMID:Recombinant Human DFF45 Inhibits Apoptosis-specific Endonuclease in a Cell-free System of Xenopus Egg Extracts. 1205 94

With a 0.5 kb probe of CX2, distribution of CX2 tandem repeats was studied in different C.albicans strains. Results suggest that all the C.albicans strains tested contained the tandem repeat. In order to verify if the expression of CX2 was hyphal specific, its expression was analyzed under various inductive and non-inductive conditions. With CX2 0.5 kb probe, Northern hybridization analysis confirmed that it was specifically in hyphae. The result of chromosomal localizationtion and genomic Southern blot analysis suggested that there were other genes containing the tandem repeat besides of ALS1. A C.albicans s genomic DNA library was screened with the CX2 0.5 kb probe and several positive recombinant lambda phages were obtained. After endonuclease identification, subcloning, and sequence analysis, several ALS family genes were cloned. No.1 lambda phage DNA contained ALS4, No.4 lambda phage DNA contained ALS1, No.6 lambda phage DNA contained ALS3. To study the role of ALS family genes in yeast-hyphal transition, als1/ALS1 mutant was constructed by homologous recombination. The ability to form hyphae was tested in different inductive culturing conditions. Defective hyphal growth were observed in some solid media.
Sheng Wu Hua Xue Yu Sheng Wu Wu Li Xue Bao (Shanghai) 2000
PMID:Cloning and Functional Analysis of ALS Family Genes from Candida albicans. 1205 69


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