Gene/Protein Disease Symptom Drug Enzyme Compound
Pivot Concepts:   Target Concepts:
Query: EC:3.1.30.2 (endonuclease)
 18,621 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

Citrullinemia is an inborn error of metabolism due to deficiency of the urea cycle enzyme, argininosuccinate synthetase [L-citrulline:L-aspartate ligase (AMP-forming), EC 6.3.4.5]. The disease was first described in humans but was recently reported in dairy cattle in Australia. Here we report the nucleotide sequence of the normal bovine cDNA for argininosuccinate synthetase and the mutation present in animals with citrullinemia. Analysis of DNA from affected animals by Southern blotting did not readily identify the mutation in the bovine gene. RNA (Northern) blotting revealed a major reduction in the steady-state amount of mRNA in the liver of affected animals to less than 5% of controls. The bovine cDNA was cloned and sequenced and revealed 96% identity with the deduced human sequence at the amino acid level. Starting with mutant bovine liver, the mRNA was reverse-transcribed; the cDNA product was amplified with the polymerase chain reaction, cloned, and sequenced. The sequence revealed a C----T transition converting arginine-86 (CGA) to a nonsense codon (TGA). A second C----T transition represented a polymorphism in proline-175 (CCC----CCT). The mutation and the polymorphism were confirmed by amplification of genomic DNA and demonstration with restriction endonuclease enzymes of both the loss of an Ava II site in DNA from mutant animals at codon 86 and the presence or absence of a Dde I site at codon 175. The loss of the Ava II site can be used for rapid, economical, nonradioactive detection of heterozygotes for bovine citrullinemia.
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PMID:Molecular definition of bovine argininosuccinate synthetase deficiency. 281 70

Specific endonuclease activities have been found it two Pseudomonas aeruginosa strains. Isolation and purification of enzymes and determining their specific activities have permitted one to find out that PaeI is an isoshizomer of SphI and digests the sequence 5'-GCATG C-3'. Another isolated enzyme PaeII is an isoshizomer of SmaI and cleaves DNA in a fragment 5'-CCC GGG-3'. The use of PaeI and PaeII enzymes in genetical engineering and their advantages are discussed.
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PMID:[New specific endonucleases PaeI and PaeII from Pseudomonas aeruginosa]. 302 9

Restriction of the covalently closed circular DNA from phage PM2 (CCC PM2 DNA) by Hap II endonuclease was studied under varying enzyme concentration. At low Hap II concentration, accumulation of the intermediate product, OC DNA, was observed at the early stages of the reaction. The resulting final mixture of restriction products consists of OC and L DNA, and their relative content depends on the concentration of the enzyme used. The affinity of the enzyme for the intact recognition site of the substrate in different conformational forms does not seem to be affected. Basically identical results were obtained with the two different CCC DNA used: PM2 and SV40 DNA.
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PMID:Restriction of PM2 supercoiled DNA by Hap II endonuclease. 626 36

Based on the results of a systematic study of factors affecting plasmid yield and purity, a procedure suitable for the rapid screening for and isolation of covalently closed circular DNA from Streptomyces lividans and Escherichia coli was developed. The method consists of lysis of lysozyme-treated bacteria combined with alkaline denaturation of DNA at high temperature. Renaturation of CCC DNA and precipitation of single-stranded DNA together with protein is achieved by the addition of a minimal amount of phenol/chloroform. The screening procedure uses only a single tube and the samples can be analyzed by agarose gel electrophoresis about 30 min after lysis. Removal of phenol and further purification of the plasmid preparation is achieved by consecutive precipitations with isopropanol and spermine, followed by extraction with ethanol, producing samples suitable for restriction endonuclease digestion, ligation, and transformation of S. lividans protoplasts or competent E. coli cells in about 2 h. All steps of the procedure are explained in detail with information about the effects of changing parameters. This should help the experimenter to obtain reproducible results and may be useful if the method has to be adapted to new strains or plasmids.
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PMID:Factors affecting the isolation of CCC DNA from Streptomyces lividans and Escherichia coli. 638 33

The XmaI endonuclease recognizes and cleaves the sequence C decreases CCGGG. Magnesium is required for catalysis, however, the enzyme forms stable, specific complexes with DNA in the absence of magnesium. An association constant of 1.2 x 10(9)/M was estimated for the affinity of the enzyme for a specific 195 bp fragment. Competition assays revealed that the site-specific association constant represented an approximately 10(4)-fold increase in affinity over that for non-cognate sites. Missing nucleoside analyses suggested an interaction of the enzyme with each of the cytosines and guanines within the recognition site. Recognition of each of the guanines was also indicated by dimethylsulfate interference footprinting assays. The phosphates 5' to the guanines within the recognition site appeared to be the major sites of interaction of XmaI with the sugar-phosphate backbone. No significant interaction of the protein was observed with phosphates flanking the recognition sequence. Comparison of the footprinting patterns of XmaI with those of the neoschizomer SmaI (CCC decreases GGG) revealed that the two enzymes utilize the same DNA determinants in their specific interaction with the CCCGGG recognition site.
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PMID:DNA determinants in sequence-specific recognition by XmaI endonuclease. 756 71

SmaI endonuclease recognizes and cleaves the sequence CCC decreases GGG. The enzyme requires magnesium for catalysis; however, equilibrium binding assays revealed that the enzyme binds specifically to DNA in the absence of magnesium. A specific association constant of 0.9 x 10(8) M-1 was determined for SmaI binding to a 22-base duplex oligonucleotide. Furthermore, the KA was a function of the length of the DNA substrate and the enzyme exhibited an affinity of 1.2 x 10(9) M-1 for a 195-base pair fragment and which represented a 10(4)-fold increase in affinity over binding to nonspecific sequences. A Km of 17.5 nM was estimated from kinetic assays based on cleavage of the 22-base oligonucleotide and is not significantly different from the KD estimated from the thermodynamic analyses. Footprinting (dimethyl sulfate and missing nucleoside) analyses revealed that SmaI interacts with each of the base pairs within the recognition sequence. Ethylation interference assays suggested that the protein contacts three adjacent phosphates on each strand of the recognition sequence. Significantly, a predicted protein contact with the phosphate 3' of the scissile bond may have implications in the mechanism of catalysis by SmaI.
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PMID:Sequence-specific DNA recognition by the SmaI endonuclease. 789 84

We used a polymerase chain reaction (PCR) strategy and restriction fragment polymorphism analysis to evaluate all 19 exons of the plasminogen (PLG) gene in a Japanese patient with congenital PLG deficiency and her family members. She presented with cerebral infarction. Sequence analysis following amplification of each exon and its flanking regions showed a single T to C transition in exon 14, which changed a Ser572 codon (TCC) to Pro572 codon (CCC). Since this mutation generates a new Fok I site, the Fok I digestion pattern of the PCR-amplified exon 14 fragments from each family member was analyzed. In all cases, the patterns were consistent with the activities and antigen levels of plasma PLG in those members. Furthermore, all PCR-amplified exon 14 fragments from 15 normal individuals were not restricted with Fok I endonuclease. We conclude that a T to C transition in exon 14 identified in the propositus is responsible for PLG deficiency inherited in this Japanese family with thrombotic episodes.
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PMID:Congenital plasminogen deficiency caused by a Ser572 to Pro mutation. 839 98

Acoording to the telomere-repeated sequences of rice, two primers: (TTTAGGG11)(3) and (CCC-TAA.A)(3)CCC were used to amplify rice telomere-assciated sequences (TASs). Fox PCR template preparation. total DNA was digested with restrictive endonuclease and then ligated. Using the ligates or total DNA sa template, eight fragments were obtained with the single primer by the PCR reaction. To confirm that the sequences are derived from telomeric DNA, we conducted Bal31 digestion analysis. Of the eight fragments, seven were susceptible to Bal31 treatment, suggesting that they were TASs. These DNA fragments were further demonstrated u, be rice sub-telomeric sequences by RFLP mapping Five sequences have been mapped to the distal ends on rice chromme 5,6,7 and 9, and two other sequences have been mapped at interstitial sites, suggesting that (TTTAGGG)(n). also exist in the middle of rice chromosomes-All eight fragments were sequenced and characterized.
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PMID:Isolation and characterization of five rice telomere-associated sequences. 1872 53