Gene/Protein Disease Symptom Drug Enzyme Compound
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Query: EC:3.1.30.2 (endonuclease)
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DNA fingerprinting of 15 reference strains and 24 clinical isolates of Chlamydia trachomatis, 2 strains of C. psittaci and one strain of C. pneumoniae was studied by use of universal 16 + 23S RNA from Escherichia coli, 16S rDNA-directed oligonucleotide and randomly cloned chlamydial DNA probes. The rRNA-gene restriction patterns (ribotypes) enabled the differentiation of chlamydial species. Following DNA cleavage by restriction endonuclease PvuII, lymphogranuloma venereum and trachoma biovars of C. trachomatis could be differentiated. An oligonucleotide, designed to hybridize the C. trachomatis 16S rDNA, also allowed for both species-specific identification and biovar typing of C. trachomatis human strains. Molecular typing system using 3 lambda clones containing C. trachomatis serotype E random DNA inserts, combined to ribotyping, revealed 12 groups of variable banding patterns within C. trachomatis, and could provide an alternative epidemiological tool.
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PMID:DNA fingerprinting of Chlamydia trachomatis by use of ribosomal RNA, oligonucleotide and randomly cloned DNA probes. 129 28

Twelve serovar type strains (C, D, E, F, G, H, I, J, K, L1, L2 and L3) and eight isolates (D, 4; E, 2; K, 2) of C. trachomatis were examined by restriction endonuclease analysis (REA) of DNA extracted from the elementary bodies. No difference was observed in DNA patterns of three serotypes (L1, L2 and L3) of the biovar LGV when they were digested with EcoRI and analysed by electrophoresis in a 0.6% agarose gel. The genital strains of the biovar trachoma (serovars D-K) showed similar EcoRI patterns with or without detectable differences. Serovar C of the biovar trachoma differed from the biovar LGV and the genital strains. Comparative analysis of DNAs digested with EcoRI, BamHI, HindIII, SalI and NcoI revealed that C. trachomatis isolates belonging to serovars D and K, but not E, could be subdivided into different genome types. These results suggest that DNA cleavage pattern analysis is useful for epidemiological, clinical, and taxonomic studies of C. trachomatis.
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PMID:[Restriction endonuclease analysis of Chlamydia trachomatis DNA]. 168 Sep 37

The 60-kDa cysteine-rich outer membrane protein genes of Chlamydia psittaci, Chlamydia pneumoniae, and Chlamydia trachomatis have very different 5' ends, but two areas flanking this variable region show absolute sequence conservation. This observation permitted differentiation of the three species of Chlamydia by the polymerase chain reaction (PCR), forming the basis of a diagnostic test for chlamydial infections. The PCR product containing the variable region of the respective 60-kDa CrP genes was also subjected to restriction endonuclease digestion, enabling differentiation of individual type strains of C. psittaci. Differentiation was possible between lymphogranuloma venereum and trachoma isolates of C. trachomatis. The PCR-based diagnostic test was successful with all strains of chlamydiae studied. The PCR primers showed high specificity and did not produce any product with common bacterial pathogens that may share the same sites of infection.
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PMID:Genetic diversity and identification of human infection by amplification of the chlamydial 60-kilodalton cysteine-rich outer membrane protein gene. 186 38

DNAs from eight Chlamydia psittaci isolates (koala conjunctivitis, avian psittacosis, avian ornithosis, ovine abortion, ovine polyarthritis, sporadic bovine encephalomyelitis, and feline conjunctivitis) and one Chlamydia trachomatis isolate (lymphogranuloma venereum) were compared by restriction endonuclease and DNA probe analyses. Digestion with HindIII yielded a series of discrete fragments which allowed the differentiation of most isolates. A gene probe, pFEN207, which encodes the chlamydia-specific component of the lipopolysaccharide group antigen was used in Southern hybridizations. The probe was chlamydia specific and hybridized to a single BamHI fragment and multiple HindIII fragments in each isolate. The variation in size of the hybridizing fragments allowed easy differentiation of the isolates and may eventually lead to a meaningful subgrouping of the diverse group of disease agents presently included in the species C. psittaci.
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PMID:Comparison of Chlamydia psittaci isolates by restriction endonuclease and DNA probe analyses. 282 36

DNA from a total of 60 Chlamydia trachomatis isolates was examined by restriction endonuclease analysis. Strains from all established biovars and serovars were tested. There was great diversity between the mouse biovar and the lymphogranuloma venereum (LGV) and trachoma biovars. The LGV and trachoma biovar isolates generated similar fragment patterns; however, distinct fragments appeared to be unique to both biovars, thus allowing differentiation of these two major groups. In most cases, strains of the same serovar could be differentiated from one another when a battery of restriction enzymes was used. In addition, in some cases, certain restriction fragments appeared to be characteristic of strains from a particular geographical location. The DNA patterns generated by all C. trachomatis isolates differed greatly from the DNA patterns generated from the Chlamydia psittaci isolates tested, including TWAR, a human C. psittaci strain.
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PMID:Restriction endonuclease analysis of DNA from Chlamydia trachomatis biovars. 283 86

Plasmids from Chlamydia trachomatis LGV-434 (serotype L2) and Chlamydia psittaci meningopneumonitis strain Cal-10 were cloned into the BamHI and EcoRI sites of pBR322, respectively. The recombinant plasmids pCTL2 and pCPMn, each containing an entire respective chlamydial plasmid, were transformed into Escherichia coli. The sizes of the plasmids of C. trachomatis and C. psittaci were 7.3 and 6.2 kilobases, respectively. The two plasmids were found to be distinct by restriction endonuclease analysis, DNA-DNA hybridization, and electron microscopic heteroduplex analysis. However, partial homology was observed between restriction fragments of pCTL2 and pCPMn by Southern blot analysis. Polypeptide products encoded by these plasmids were synthesized in vitro by an E. coli-directed transcription-translation system and in vivo in E. coli maxicells and minicells. None of these polypeptides was immunoreactive with anti-chlamydial sera by immunoblotting or immunoprecipitation. Based on the comparative analysis data, the C. trachomatis and C. psittaci plasmids were found to share little genetic relatedness.
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PMID:Molecular characterization of Chlamydia trachomatis and Chlamydia psittaci plasmids. 394 8

The DNA from six serovars of Chlamydia trachomatis, lymphogranuloma venereum (LGV) I, LGV II, LGV III, B, C, and D, and from Chlamydia psittaci was extracted, treated with restriction endonuclease enzymes, and run on agarose gels. By using this technique, the DNA of C. trachomatis could be clearly differentiated from C. psittaci DNA. A comparison of the DNA from the different serovars of C. trachomatis revealed similar patterns with and without detectable differences. LGV I, LGV II, LGV III, B, and C revealed no differences when treated with BamHI, HaeIII, XbaI, and XhoI. LGV III DNA, when cleaved with EcoRI and HhaI, had a major band migrating faster than the other two LGV serovars. Serovar D had a different pattern from all other strains tested when cleaved with BamHI, EcoRI, HhaI, HincI, and XhoI. When treated with SacI and HgaI, LGV II displayed a unique band not seen in the other LGV serovars. Differences in strains could be attributed to both chromosomal and plasmid DNA.
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PMID:Characterization of Chlamydia DNA by restriction endonuclease cleavage. 630 76

We have used nested polymerase chain reaction (PCR) and the PCR-based endonuclease digestion method to genotype Chlamydia trachomatis serovars in 460 infected individuals from the Eastern Highlands Province of Papua New Guinea. Our study groups comprised women who presented in labour to the Goroka Base Hospital, their newborn infants, symptomatic children who presented to the hospital's Outpatients Department and men and women from 15 randomly selected villages in the Asaro Valley. In this analysis, the major outer membrane protein (MOMP) gene, omp1, of C. trachomatis was amplified using DNA obtained from the endocervix of women, urine from men, and both the eye and nasopharynx of children. Amplified DNAs were digested concurrently using Alul and a combination of EcoRI, Hinl and Hpall restriction enzymes. The mixtures were separated on electrophoretic gels and the respective serovars designated on the basis of resolved digested DNA patterns. Our results, which were confirmed also by omp1 sequence data, show serovars D, E, F, G, H and L3 to be present in the studied communities. The overall relative frequencies of these serovars were 30%, 21%, 25%, 1%, 20% and 2% respectively, with serovars D, E, F and H accounting for 97% of these infections. Double infections among these principal serovars were also detected in all our study groups but at a low overall frequency of 3%. Serovar D was the major agent involved in the aetiology of chlamydial infection in both children and adults though serovar F was the most frequent in newborn infants. Serovar H was relatively less frequent in symptomatic children. No trachoma-related serovars were detected, confirming the rarity of this disease in Papua New Guinea. In contrast, although clinical cases of lymphogranuloma venereum have not been described in the country, the detection of serovar L3 in this study suggests that it may occur. However, the association of L3 also with childhood infection indicates that it may be causing the same pathology as the serovars D-K that are associated with non-ulcerative sexually transmitted infections.
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PMID:Chlamydia trachomatis infection and distribution of serovars in the Eastern Highlands Province, Papua New Guinea. 1958 96