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)

To examine the role of the alpha 2-adrenoceptor gene in the development of genetic hypertensive rats, we tested Dahl salt sensitive (S) and resistant rats (R) for the presence of a restriction fragment length polymorphism (RFLP) in that gene. An RFLP was found between the S and R rats with a human kidney cDNA alpha 2-adrenoceptor probe (alpha 2-C4) and Msp I restriction endonuclease. The alpha 2-C4 probe detected two alleles, S and R, of 3.0 and 2.8 kb in size. The two strains of rats were each homozygous for their corresponding allele. The inheritance of the alleles was investigated by crossbreeding S and R rats and subsequent brother/sister mating of F1 rats. Two hundred and fifteen F2 rats were produced by breeding 14 pairs of F1 rats. An atypical distribution of alpha 2-adrenoceptor genotypes was observed. We found a reduced number of the SS genotype in both males and females of the F2 generation. To investigate the mechanism of this distorted alpha 2-adrenoceptor genotype distribution in the F2 rats of S and R cross, we backcrossed the F1 rats to their S and R parents. The litter size and gender distribution were counted for each breeding colony. Analysis by chi-square test showed that there was no sex difference among the backcrosses. Also, there was no significant decrease in litter size. This excludes the possibilities of fetal demise of S homozygotes and intrauterine selection. Therefore the deficiency of SS genotype may be due to gene recombination or may not be due to the alpha 2-adrenoceptor gene itself, but to the effect of other genes closely linked to the alpha 2-adrenoceptor.
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PMID:The distorted alpha 2-adrenoceptor genotype distribution in F2 populations of Dahl salt sensitive and resistant rats cross. 810 Sep 79

The macrovascular complications of non-insulin-dependent diabetes mellitus (NIDDM) are related to the features of insulin resistance (IR). High Factor VII:C (FVII:C) levels are associated with increased cardiovascular risk and relate to a base change in the FVII gene detected by Msp I endonuclease, and also to an insertion polymorphism in the promoter region. To examine the association between FVII:C levels, genotype and features of IR, 95 NIDDM patients were studied. Genotype was related to FVII:C levels (M1M1 137%, n = 75; M1M2 and M2M2 114%, n = 20, p < 0.005; AA 136%, n = 71; Aa 119%, n = 21, p < 0.05), which is consistent with previous studies in healthy populations. FVII:C correlated with cholesterol (r = 0.51, p < 0.0005), insulin (r = 0.36, p = 0.002), triglycerides (r = 0.34, p = 0.001), age (r = 0.23, p < 0.005) and body mass index (r = 0.23, p < 0.05). When analysed by Msp I genotype, the stronger predictor of FVII:C levels, these correlations remained, with no difference in regression slopes. In a multiple regression model, genotype, cholesterol, insulin, and gender remained as independent predictors of FVII:C levels. In conclusion, FVII:C concentrations are elevated in NIDDM in relation to both FVII genotypes and features of IR.
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PMID:Factor VII gene polymorphisms, factor VII:C levels and features of insulin resistance in non-insulin-dependent diabetes mellitus. 870 97

The regulation of the Sso II restriction-modification system from Shigella sonnei was studied in vivo and in vitro . In lacZ fusion experiments, Sso II methyltransferase (M. Sso II) was found to repress its own synthesis but stimulate expression of the cognate restriction endonuclease (ENase). The N-terminal 72 amino acids of M. Sso II, predicted to form a helix-turn-helix (HTH) motif, was found to be responsible for the specific DNA-binding and regulatory function of M. Sso II. Similar HTH motifs are predicted in the N-terminus of a number of 5-methylcytosine methyltransferases, particularly M. Eco RII, M.dcm and M. Msp I, of which the ability to regulate autogenously has been proposed. In vitro, the binding of M. Sso II to its target DNA was investigated using a mobility shift assay. M. Sso II forms a specific and stable complex with a 140 bp DNA fragment containing the promoter region of Sso II R-M system. The dissociation constant (Kd) was determined to be 1.5x10(-8) M. DNaseI footprinting experiments demonstrated that M. Sso II protects a 48-52 bp region immediately upstream of the M. Sso II coding sequence which includes the predicted -10 promoter sequence of M. Sso II and the -10 and -35 sequences of R. Sso II.
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PMID:Specific binding of sso II DNA methyltransferase to its promoter region provides the regulation of sso II restriction-modification gene expression. 915 10

A high sensitivity method for detecting low level mutations is under development. A PCR reaction is performed in which a restriction site is introduced in wild-type DNA by alteration of specific bases. Digestion of wild-type DNA by the cognate restriction endonuclease (RE) enriches for products with mutations within the recognition site. After reamplification, mutations are identified by a ligation detection reaction (LDR). This PCR/RE/LDR assay was initially used to detect PCR error in known wild-type samples. PCR error was measured in low |Deltap K a| buffers containing tricine, EPPS and citrate, as well as otherwise identical buffers containing Tris. PCR conditions were optimized to minimize PCR error using perfect match primers at the Msp I site in the p53 tumor suppressor gene at codon 248. However, since mutations do not always occur within pre-existing restriction sites, a generalized PCR/RE/LDR method requires the introduction of a new restriction site. In principle, PCR with mismatch primers can alter specific bases in a sequence and generate a new restriction site. However, extension from 3' mismatch primers may generate misextension products. We tested conversion of the Msp I (CCGG) site to a Taq I site (TCGA). Conversion was unsuccessful using a natural base T mismatch primer set. Conversion was successful when modified primers containing the 6 H,8 H -3, 4-dihydropyrimido[4,5- c ][1,2]oxazine-7-one (Q6) base at 3'-ends were used in three cycles of preconversion PCR prior to conversion PCR using the 3' natural base T primers. The ability of the pyrimidine analog Q6 to access both a T-like and C-like tautomer appears to greatly facilitate the conversion.
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PMID:Nucleotide analogs and new buffers improve a generalized method to enrich for low abundance mutations. 1010 Nov 89

The genetic homogeneity of 37 strains of ruminal streptococci was investigated by comparing DNA fragment profiles on agarose gels following restriction endonuclease digestion with Hae III, Cfo I and Msp I. Thirty strains were indistinguishable from Streptococcus bovis strains, 2B, H24 and AR3. The remaining three strains were similar but not identical to a ruminal strain of Strep. intermedius (AR36). In addition, the susceptibility of these strains to infection by five bacteriophages was examined. Three of the phages (phi Sb02, phi Sb03 and phi Sb04) were specific to the strain of Strep. bovis from which they were isolated, while phages 2BV and phi Sb01 infected one and two strains, respectively, in addition to their primary host. It was concluded that although Strep. bovis is relatively homogeneous genetically, broad host range phages appear to be uncommon with this bacterial species.
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PMID:Genetic homogeneity and phage susceptibility of ruminal strains of Streptococcus bovis isolated in Australia. 1049 98

We examined the possible association between a transforming growth factor (TGF)-beta(1) gene polymorphism in codon 10 and blood pressure (BP) at rest, in acute response to exercise in the pretrained (sedentary) and trained states, as well as in its training response (Delta) to 20 wk of endurance exercise. Subjects were 257 black and 480 white, healthy sedentary normotensive subjects from the HERITAGE Family Study. The polymorphism was detected by polymerase chain reaction and digestion with the Msp A1 I endonuclease yielding a wild (leucine-10) and a mutant (proline-10) allele. Resting and exercise [50 W plus 60, 80, and 100% maximal oxygen consumption (VO(2)(max))] BP were determined before and after training. Significant (P < 0.05) race-genotype interactions were found for systolic (S) BP in both the sedentary and trained states. Among whites but not in blacks, the TGF-beta(1) genotypes were significantly (P < 0.05) associated with sedentary-state SBP at rest, at 50 W, and at 60 and 100% VO(2)(max)as well as with trained-state SBP at rest and at 80 and 100% VO(2)(max). The leucine-10 homozygotes had significantly (P < 0.05) lower SBP than proline-10 homozygotes. DeltaBP was not significantly associated with genotype. These results support the hypothesis of an association between the TGF-beta(1) marker in codon 10 and SBP at rest and in response to acute exercise in whites but not in blacks.
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PMID:TGF-beta(1) gene-race interactions for resting and exercise blood pressure in the HERITAGE Family Study. 1156 66

By means of in situ nick-translation technique, methylation patterns of pericentric heterochromatin of chromosomes 1, 9 and 16 in extraembryonic (chorion) and embryonic cells of 5-8 week old human fetuses with normal karyotype (5), and in one specimen with trisomy for chromosome 16 were studied. Fixed metaphase chromosomes from direct chromosome preparations were digested with either endonuclease Msp I or its isoshizomer Hpa II recognizing and restricting the same sDNA sequence C decreases CGC with Hpa II, but not Msp I sensitive to methylation state of internal cytosin. According to our results, heterochromatin of extraembryonic, but not embryonic cells is hypomethylated. An obvious difference was registered in signal strength between homologous regions in iq12 of both parental chromosomes 1 in early (5-6 week old), but not in more advanced fetuses. Methylation pattern difference was detected in pericentric chromatin of triple copies of chromosome 16 in extraembryonic tissues of the 47,XY, + 16 fetus. These results are in line with a hypothesis of intraheterochromatin location of "early" genes governing initial stages of embryonic development in humans.
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PMID:[The methylation peculiarities of pericentromeric heterochromatin of chromosomes 1,9 and 16 in human embryo]. 1160 93

Toll-like receptor 4 recognizes pathogen ligands and mediates signaling to initiate innate and adaptive immune responses. In this experiment, a 477 bp segment of the 5'-flanking region of TLR4 gene of Chinese Holstein, Sanhe cattle and Chinese simmental was amplified by polymerase chain reaction. After sequencing, a polymorphic site in amplified production of TLR4 was identified of having either a G or a C at position 245. This polymorphism in the three populations was detected by digesting the fragment with restriction endonuclease Msp I. Results showed that both alleles (A and B) were found in the three populations and the value of polymorphism information content indicated that this was a moderate polymorphism. Chi2 test indicated that the polymorphism locus in Sanhe cattle did not fit Hardy-Weinberg equilibrium (P< 0.05). In addition, the effect of the TLR4 polymorphism on somatic cell score was analyzed, and the results indicated that the somatic cell score were significantly affected by lactation month and the type of breeds(P<0.05), but not by different genotypes (P > 0.05).
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PMID:[Genetic variation in the 5' flanking region of bovine TLR4 gene and correlation with mastitis]. 1713 37

For a better understanding of epigenetic regulation of cell differentiation, it is important to analyze DNA methylation at a specific site. Although previous studies described methylation of isolated DNA extracted from cells and tissues using a combination of appropriate restriction endonucleases, no application to tissue cell level has been reported. Here, we report a new method, named histo endonuclease-linked detection of methylation sites of DNA (HELMET), designed to detect methylation sites of DNA with a specific sequences in a tissue section. In this study, we examined changes in the methylation level of CCGG sites during spermatogenesis in paraffin-embedded sections of mouse testis. In principle, the 3'-OH ends of DNA strand breaks in a section were firstly labeled with a mixture of dideoxynucleotides by terminal deoxynucleotidyl transferase (TdT), not to be further elongated by TdT. Then the section was digested with Hpa II, resulting in cutting the center portion of non-methylated CCGG. The cutting sites were labeled with biotin-16-dUTP by TdT. Next, the section was treated with Msp I, which can cut the CCGG sequence irrespective of the presence or absence of methylation of the second cytosine, and the cutting sites were labeled with digoxigenin-11-dUTP by TdT. Finally, both biotin and digoxigenin were visualized by enzyme- or fluorescence-immunohistochemistry. Using this method, we found hypermethylation of CCGG sites in most of the germ cells although non-methylated CCGG were colocalized in elongated spermatids. Interestingly, some TUNEL-positive germ cells, which are frequent in mammalian spermatogenesis, became markedly Hpa II-reactive, indicating that the CCGG sites may be demethylated during apoptosis.
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PMID:In situ detection of methylated DNA by histo endonuclease-linked detection of methylated DNA sites: a new principle of analysis of DNA methylation. 1879 17

Restriction endonuclease digests of total DNA from races 3, 4, and 5 of the soybean cyst nematode, Heterodera glycines, have been analyzed on agarose gels. DNA fragment patterns of race 4 were completely different from those patterns obtained for races 3 and 5 by all eight restriction enzymes tested. Differences in long and short restriction DNA fragments generated by the enzyme Msp I or its isoschizomer, Hpa II, were detected between race 3 and 5 digestion profiles. Rapid DNA isolation followed by its digestion with either Msp I or Hpa II enzymes and visualization of repetitive DNA fragments in agarose gels provided a diagnostic assay for the populations of the three races examined in this study.
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PMID:DNA Restriction Fragment Length Polymorphism in Races of the Soybean Cyst Nematode, Heterodera glycines. 1929 Feb 51


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