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)

We have previously identified a unique site, pac, from which packaging of precursor concatameric viral DNA into proheads starts during the maturation process of bacteriophage CP-T1. The direction of this packaging was determined from restriction enzyme cleavage patterns of CP-T1 DNA. A restriction enzyme generated fragment containing pac was cloned and the surrounding DNA region sequenced. Analysis of the nucleotide sequence revealed numerous repeat regions related to the consensus sequence PuagttGAT.AAT.aa.t. Within the sequenced region an open reading frame encoding a 12260 Mr protein was also identified. This protein appears to share homology with the binding domains of known DNA binding proteins and may represent a putative Pac terminase possessing the specific endonuclease activity required for cleavage at the pac site. Minicell analysis of deletion derivatives of the pac-containing clone revealed a protein of approximately 12,900 Mr encoded within this same region, confirming that this Pac protein is phage encoded.
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PMID:Molecular analysis of the packaging signal in bacteriophage CP-T1 of Vibrio cholerae. 341 20

The amidase genes of Pseudomonas aeruginosa were inserted into a lambda replacement vector following cleavage with the restriction endonuclease HindIII. The recombinant lambdaami was detected by enhanced growth of Escherichia coli around plaques of the recombinant phage on minimal medium containing acetamide as the nitrogen source. Low levels of amidase activity were detected in E. coli cultures infected with lambdaami and these were sufficient to allow growth with acetamide as nitrogen source. Lysis-defective derivatives of lambdaami were made by introducing Q-, S-, mutations. Cultures of E. coli infected with lambdaamiQ-S- synthesised amidase as the major protein. The amidase produced by these cultures was identical to that produced by PAC strains of P. aeruginosa in substrate specificty, thermal stability and immunological cross-reaction.
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PMID:The construction in vitro of derivatives of bacteriophage lambda carrying the amidase genes of Pseudomonas aeruginosa. 624 42

We studied the DNA sequence of the entire coding region of ERCC1 gene, in five cell lines established from human ovarian cancer (A2780, A2780/CP70, MCAS, OVCAR-3, SK-OV-3), 29 human ovarian cancer tumor tissue specimens, one human T-lymphocyte cell line (H9), and non-malignant human ovary tissue (NHO). Samples were assayed by PCR-SSCP and DNA sequence analyses. A silent mutation at codon 118 (site for restriction endonuclease MaeII) in exon 4 of the gene was detected in MCAS, OVCAR-3 and SK-OV-3 cells, and NHO. This mutation was a C-->T transition, that codes for the same amino acid: asparagine. This transition converts a common codon usage (AAC) to an infrequent codon usage (AAT), whereas frequency of use is reduced two-fold. This base change was associated with a detectable band shift on SSCP analysis. For the 29 ovarian cancer specimens, the same base change was observed in 15 tumor samples and was associated with the same band shift in exon 4. Cells and tumor tissue specimens that did not contain the C-->T transition, did not show the band shift in exon 4. Our data suggest that this alteration at codon 118 within the ERCC1 gene, may exist in platinum-sensitive and platinum-resistant ovarian cancer tissues.
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PMID:A nucleotide polymorphism in ERCC1 in human ovarian cancer cell lines and tumor tissues. 936 Jun 34

A new AAT allele (PI Zbristol) has been discovered in a woman with an obstetric history of three perinatal deaths from fulminant liver disease and no living offspring. She and her father were both PI M1Zbristol heterozygotes. The Zbristol protein is active as a proteinase inhibitor but appeared to be deficient in the plasma to about the same degree as the S protein in MS heterozygotes. It focuses on the basic side of Z and lacks the normal pattern of secondary isoforms associated with the commonly occurring AAT variants and migrates faster than normal on an SDS electrophoresis gel. The Zbristol mutation was found to be a C to T transition at codon 85 changing ACG (Thr) to ATG (Met). This disrupts the N-glycosylation site starting at Asn 83 preventing glycosylation at residue 83 in the PI Zbristol protein and explains the protein isoelectric focusing and SDS gel electrophoresis results. An analysis of haplotypes in the propositus and her father indicated that the Zbristol mutation occurred on the common M1(Val 213) genetic background. The new mutation also led to the generation of an NlaIII restriction endonuclease recognition site. Cell lines from two offspring tested for the presence of this NlaIII site revealed that one had the variant and the other did not. Thus, the relationship between Zbristol and fulminant liver disease in the offspring is unclear.
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PMID:A new alpha 1-antitrypsin mutation, Thr-Met 85, (PI Zbristol) associated with novel electrophoretic properties. 945

The Schizosaccharmyces pombe pac-1 gene product is a kind of dsRNA dependent ribonuclease, which has potential to degrade the dsRNA viral genome, the replication form of ssRNA viral genome and viroid genome. Therefore, to introduce the pac-1 gene into plants conferring them resistance to viruses is a new method of establishing the anti-virus transgenic plant. The pac-1 gene from the S. pmobe genome DNA isolated from China was cloned by means of PCR amplification. The pac-1 gene was inserted into the cloning vector pGEM-7Zf(+) by using restriction endonuclease Kpn I/BamHI. Sequencing analysis shows that it is a complete gene with 1095 necleotides. Compared to the reported pac-1 gene, its homology is significant, but with 5 nucleotides differences, leading to only one amino acid difference. Pac-1 gene was inserted into the prodaryotic expression vector pET-21(a) by using the restriction endonuclase Nde I/BamHI. It was induced by the IPTG in E. coli BL21 harbouring the recombinant vector pET-pac-1. The pac-1 gene product is analyzed by the SDS-PAGE. The result shows the product of pac-1 gene exists in the supernatant part as soluble form and in the precipitant part as inclusion bodies after the cells were lysed by ultrasonic wave. The supernatant was applied to detect the enzyme activity of pac-1 gene product. We concluded that pac-1 gene has the biological activity of degrading the CMV-dsRNA.
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PMID:[Cloning, sequence analysis and high-level expression in Escherichia coli and activity assay of pac-1 gene from Schizosaccharmyces pombe]. 1141 Dec 32

Human rotavirus is the major etiologic agent of infantile diarrhea on a worldwide scale. In this study, rotaviruses were detected by reverse-transcription PCR in 42 of 83 stool specimens from children below the age of 3 years with acute diarrhea in Bangkok, Thailand, between November 1998 and August 1999. G and P types of all samples were characterized by restriction endonuclease analysis (REA) and multiplex PCR typing assay, respectively. Strain G1P[8] (76.1%) was the predominant type, followed by G1P[6] (2.4%). Strain G1 combined with mixed P[8]/P[6] was identified in 2 specimens (4.8%) and 7 untypeable G strains (16.7%) were observed. This information on the circulating G and P combinations should be useful for understanding the epidemiology of human rotavirus in Bangkok, Thailand.
Asian Pac J Allergy Immunol 2001 Mar
PMID:Predominant human rotavirus genotype G1P[8] infection in infants and children in Bangkok, Thailand. 1149

Eight human viruses of the Herpesviridae family represent a significant public health problem world-wide. Detection and typing of five of the human herpesviruses (HSV-1, HSV-2, VZV, EBV, and CMV) was performed by applying a consensus primer polymerase chain reaction (PCR). The amplified PCR products from the five human herpesviruses were typed based on their restriction enzyme digestion polymorphism with Hinf I and Alu I. Fifteen clinically suspected specimens from herpesvirus-infected patients were also evaluated. A fragment of the DNA polymerase gene from each of the five human herpesviruses was successfully amplified by the set of consensus primers. Their amplicons obtained by PCR from the template DNAs were subjected to restriction endonuclease digestion and human herpesviruses 1-5 could be clearly differentiated and typed. This method can be used to detect and differentiate between the five human herpesviruses in clinical specimens. This study demonstrates the value of testing for five human herpesviruses by consensus PCR and restricted fragment length polymorphism (RFLP). These procedures are simple and straightforward techniques for the investigation of clinical specimens.
Asian Pac J Allergy Immunol 2003 Mar
PMID:Detection and differentiation of human herpesviruses 1-5 by consensus primer PCR and RFLP. 1293 52

Applying various restriction enzymes on a specially designed 1.5 kb DNA fragment revealed that the inhibitory effects of PUVA treatment on restriction endonuclease activities are caused by recognition inhibition. In this study, Restriction enzymes which have a 5'-TpA sequence at the cleaving site (Kpn I, Xba I, Pme I, and Dra I), and non-cleaving site (Pac I) in recognition sites, or have two 5'-TpA sequences at the recognition site and a non-specific sequence between recognition and cleaving site (BciV I) were inhibited by PUVA treatment. Most of the other restriction enzymes used in this study which do not have a 5'-TpA sequence at their restriction site were not inhibited by PUVA treatment, although a 5'-TpA sequence is located adjacent (Sma I) or very close (BamH I, Sac I and Pst I) to the recognition and cleaving site for them.
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PMID:Inhibition of restriction enzyme's DNA sequence recognition by PUVA treatment. 1451 Apr 98

In the present study we analysed the vitamin D receptor (VDR) Fokl polymorphism distribution in a Caucasian population from the North-West of Spain. This polymorphism has been associated with variation in bone mineral density levels and has been proposed as a factor for the genetic predisposition to osteoporosis. This study was performed in 60 healthy individuals by restriction fragment length polymorphism (RFLP) analysis using the endonuclease Fokl and alternatively using single strand conformation polymorphism (SSCP). The genotype distribution was: 46.7% FF homozygotes, 43.3% Ff heterozygotes, and 10% ff homozygotes. These values do not reveal significant differences in comparison with other available Caucasoid populations, but show clear differences with respect to African-American populations. The analysis of this polymorphism by SSCP reveals the existence of a novel single nucleotide polymorphism (AAT/AAC) located within the DNA binding domain of the VDR gene at position-19 codon. Thus, using a novel 109 bp hVDR gene fragment which excludes the AAT/AAC variant in the vitamin D receptor Fokl allows an easier determination by SSCP analysis of the Fokl polymorphism.
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PMID:Analysis of the vitamin D receptor Fokl polymorphism. 1512 11

The HBV X gene was amplified by PCR according to the pecob6 containing the whole fragment of adw subtype of HBV, then the fragment was inserted into the multiple clone site of pAdTrack-CMV. The linearized shuttle plasmid was homogenously recombined with AdEasy-1 in BJ5183 and the recombinant adenoviral plasmid pAd-X was generated. Then plasmid pAd-X was digested with Pac I and transfected into 293 cells for packaging and amplifying. Infection titer and rate were monitored by green fluorescent protein (GFP) expression. With restriction endonuclease analysis and PCR methods, it has been confirmed that HBV X gene was cloned into the adenovirus vector successfully. The expression of X protein in HepG2 cells was detected by Western-blot. The recombined adenovirus Ad-X was constructed successfully, which would contribute to the advanced functional study of HBV X protein.
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PMID:[Construction of the recombined adenovirus containing HBV X gene and expression in HepG2 cells]. 1823 89


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