No abstract available.
DNA Restriction Enzymes* ; DNA* ; Ribotyping* ; Staphylococcus aureus* ; Staphylococcus*

Twenty-five isolates of Pythium species were identi6ed and classified on the basis of RFLP(restriction fragment length polymorphism) of ITS(internal transcribed spacer) region in ribosomal DNA. and M-13 PCR markers. Eight restriction endonucleases were used to investigate the genetic relatedness among isolates P. graminicola and P. arrhenomanes as well as P. aphanidermatum and P. delience produced identical band patterns with all restriction endonucleases used and M-13 markers. P. myriotylum and P. catenulatum also formed tight cluster on the basis of RFLP of ribosomal DNA but produced distinct band pattern with M-13 PCR markers. No intraspecitic variations were observed with RFLP of ITS region in ribosomal DNA. Molecular analysis based on M-13 marker demonstrated that each species produced distinct band patterns. Intraspecific variation of P. ultimum and P. torulosum was observed with M-13 markers.
DNA Restriction Enzymes ; DNA, Ribosomal* ; Polymerase Chain Reaction* ; Polymorphism, Restriction Fragment Length* ; Pythium*

G and P tying of group A porcine rotaviruses (P(o)RV) from field fecal samples were performed using reversetranscriptase polymerization chain reaction (RT-PCR) and restriction fragment length polymorphism (RFLP) analysis. After amplifying full length VP7 and partial length VP4 genes, restriction endonucleases were used to digest and analyze the cutting pattern of the gene products. After analysis of digests with restriction endonucleases, seven and six RFLP types were observed for VP7 and VP4, respectively. The G typing analysis of 50 fecal samples revealed that 68% (34/50) were G4, which included G4-like (22/50); 22% (11/50) were G5; 6% (3/50) were G4 and G5 mixed types. The P typing analysis of the same fecal samples revealed that 36% (18/50) were P2B, 52% (26/50) were P9, 1 sample (2%) was a mixture of P2B and P9. Combinations of G and P types, the G4P2B and G4P9 types including G4-like accounted for 26% (13/50) and 32% (16/50), respectively. The G5P2B and G5P9 type also represented 4% (2/50) and 18% (9/50) of the samples. No G3 and G11 or other new P types were identified from the samples tested. Information on the G and P types and G/P combinations in the field fecal samples is useful for developing more effective PoRV vaccines and understanding the epidemiology of PoRV infections in the field.
DNA Restriction Enzymes ; Epidemiology ; Polymerization ; Polymers ; Polymorphism, Restriction Fragment Length* ; Rotavirus* ; Vaccines

BACKGROUND: Recent restriction enclonuclease analysis studies hsve revealed that MCV DNA can be classified into two major types, designated MCV-1 and MCV-2, by th:ir restriction enzyme cleavsge patterns. In earlier reports of MCV DNA analysis, MCV-2 was the main virus type found in genital lesions. However many recent studies cienied the relationship between virus type and anatomical distribution. OBJECTIVE: The purpose of this study was to examine the ratio of MCV-l to MCV-2 in Korean isolates of MCV DNA and the relationship between MCV subtypes and with clinical features such as anatomical location, age, sex, numiber of lesions, and atopic dermatitis. METHODS: MCV DNA extrated from 112 cases of Korean patients waa examined by restriction endonuclease analysis using Brtm HI. RESULTS: 1. MCV-1 was found in 108 of 112 (96.4%) patients and MCV-2 in of 112 (3.6%) patients. The ratio of MCV-1 to MCV-2 wss 28:1. 2. There was no significant ciprrelation between MCV subtypes and the age, sex, number of lesions, atopic dermatitis, and anatoimic loction. 3. Lesions induced by MCV-1 MCV-2 were indistinguishable on the brsis of size and form. CONCLUSION: This study showis that the ratio of MCV-1 to MCV-2 was 28:1 in Korean molluscum contagiosum patients and there was no relationship between MCV subtyies and lesional morphology or snatomical distribution.
Dermatitis, Atopic ; DNA ; DNA Restriction Enzymes ; Humans ; Molecular Epidemiology* ; Molluscum contagiosum virus* ; Molluscum Contagiosum*

To use recombinant viruses with wider host range as viral insecticides, we investigated the characteristics and pathogenicity of host range expanded recombinant viruses in insect cells. We compared host range expanded recombinant viruses, RecS-B6 and RecB-8, constructed by cotransfection of Autographa californica nuclear polyhedrosis virus (AcNPV) and Bombyx mori NPV (BmNPV), to host range expanded AcNPV, Ac-BH, by substitution of the 0.6 Kb fragment of the BmNPV helicase gene. Restriction endonuclease profiles of RecS-B6 and RecB-8 DNAs were different from those of parent viruses. Nucleotide sequence analysis of the 0.6 Kb region in the putative helicase gene of RecS-B6 and RecB-8 showed that their structures were identical to the counterpart region of BmNPV Comparison of viral replication of these recombinant viruses in Sf-21 and BmN-4 cells showed that Ac-BH, compared to wild type viruses, replicated well in BmN-4 cells but poorly in Sf-21 cells. In contrast, RecS-B6 and RecB-8 replicated relatively well in both cells compared to parent viruses. These results may imply that random genomic recombinant viruses, RecS-B6 and RecB-8, possess better potential as viral pesticides than helicase-mediated recombinant virus, Ac-BH.
Base Sequence ; Bombyx ; DNA ; DNA Restriction Enzymes ; Host Specificity* ; Humans ; Insecticides ; Insects* ; Nucleopolyhedrovirus ; Parents ; Pesticides ; Virulence*

OBJECTIVE: This study was performed to find out mitochondrial deoxyribonucleic acid mutations in preeclampsia because Mendelian models fail to explain all the patterns of inheritance in preeclampsia. METHODS: Ten preeclampsia patients and two of their related family members who have the obstetric history of preeclampsia were studied. The mitochondrial transfer ribonucleic acidleu[UUR] gene was amplified using polymerase chain reaction, cut by a restriction endonuclease (Apa , and also sequenced to see the whole gene. RESULTS: There were neither the known mutation at Nucleotide 3243 nor other mutations on the mitochondrial transfer ribonucleic acidleu[UUR] gene in these objects. CONCLUSION: It seems that the known mutation of mitochondrial transfer ribonucleic acidleu[UUR] gene is not so frequently detected in preeclampsia of South Korean, But it could not be concluded how many South Korean women with preeclampsia have the mutation.
DNA ; DNA Restriction Enzymes ; Female ; Humans ; Point Mutation* ; Polymerase Chain Reaction ; Pre-Eclampsia* ; RNA* ; Wills

Gene cloning is one of the most important and widely used technologies in molecular biology research. Generally, DNA fragment is cut with restriction enzyme, and then the product is ligated to a linearized vector with complementary sticky end or blunt end by DNA-ligase. This traditional DNA cloning method requires compatible enzyme recognition sites existing in both PCR fragment and targeted vector. Several ligase-free methods have been established to avoid the using of restriction enzyme. However, those methods are time-consuming, labor-intensive and expensive. To overcome these shortcomings, we developed an Exonuclease III based DNA cloning method that takes only 30 minutes with high cloning efficiency and significant economic advantage. Therefore, this method is suitable for large-scale gene cloning.
Cloning, Molecular ; methods ; DNA ; chemistry ; DNA Restriction Enzymes ; Exodeoxyribonucleases ; chemistry ; Genetic Vectors ; Polymerase Chain Reaction

BACKGROUND: Methicillin-resistant Staphylococcus aureus is a major etiologic agent of hospital acquired infection. Coagulase negative staphylococci (CNS) species are previously regarded as contaminants. However nowadays CNS were regarded as an important cause of bacteremia. So in this study we wanted to analyze the patterns of plasmids and antimicrobial susceptibility test of Staphylococcus species isolated from clinical specimens. METHOD: Plasmid DNA was extracted and then processed through restriction enzyme digestion for plasmid analysis of S. aureus and antimicrobial susceptibility, which was done by agar dilution method. For S. epidermidis plasmid analysis was done without enzyme digestion. RESULTS: All of MRSA have 1 to 5 plasmids. There exists 6 patterns of S. aureus plasmid without enzyme digestion. With EcoRI and HindIII digestion pattern were more distinct and clear. For S. epidermidis enzyme digestion is not needed. Antimicrobial susceptibility patterns of S. aureus are simple whereas S. epidermidis showed variable patterns. CONCLUSIONS: For the plasmid analysis of S. aureus restriction enzyme digestion is required and for the S. epidermidis, the pattern of plasmids are variable so without restriction enzyme analysis we can obtain several patterns. Plasmid analysis will be used as a good epidemilogical tool for Staphylococcus.
Agar ; Bacteremia ; Coagulase ; Digestion ; DNA ; DNA Restriction Enzymes* ; Methicillin-Resistant Staphylococcus aureus ; Plasmids* ; Restriction Mapping ; Staphylococcus aureus* ; Staphylococcus*

Characterizations of the VP4 (P type) and VP7 (G type) genes of Korean isolates of bovine rotavirus were performed using RT-PCR/RFLP and nucleotide sequencing analysis. After RT-PCR amplification of partial length (1094bp) of the VP4 and full length (1062bp) of the VP7 genes, amplified PCR products were digested with restriction endonucleases and digestion patterns were compared with those of reference rotaviruses. With the VP4 genes, four RFLP (AD) profiles were observed; three (A, B and C) were the same as those of bovine rotavirus NCDV (P[1]), IND (P[5]) and B223 (P[11]), respectively, Profile D was the same as that of porcine rotavirus OSU (p[7]). With the VP7 genes, five RFLP profiles (I-V) were observed; three of them (1, II and III) were the same as those of bovine rotavirus NCDV (G6), Cody I-801 (G8), and B223 (G10), respectively, Profile IV and V were atypical to those of reference bovine rotaviruses used in this study. These two profiles were identified as G6 and G5, respectively, after analyzing and comparing the nucleotide sequences. The G typing analysis revealed that 61.9% (26/42) were G6, which included G6 subtype; 28.6% (12/42) were G5; 7.1% (3/42) were G10; 2.4% (1/42) were G8. The P typing analysis revealed that 54.8% (23/42) were P(5); 28.6% (12/42) were P(7); 11.8% (5/42) were P(11); 4.8% (2/42) were P(1). Our results showed that G6/P(5) were the most prevalent rotaviruses in diarrheic calves in Korea. Also, this is the first report that G5P(7) rotaviruses were identified from cattle with diarrhea.
Animals ; Base Sequence ; Cattle ; Diarrhea ; Digestion ; DNA Restriction Enzymes ; Korea ; Polymerase Chain Reaction ; Polymorphism, Restriction Fragment Length* ; Rotavirus*

Acinetobacter species encounters frequently with clinical specimens and now accounts for a substantial proportion of endemic nosocomial infections in Korea. Recent trends indicate that the antimicrobial resistant strains of Acinetobacter species are increasing. Sixty-one strains were isolated from specimens of patients suspected of nosocomial infections during 1991 to 1996. At present, phenotypic identification of Acinetobacter using biochemical test may not be reliable and resulted in the difficulty to clarify the source of infections and epidemiological study of hospital-acquired infections. Aware of the importance of rational taxonomic proposal for these isolates, correct species identification of these organisms by molecular typing method was carried out. A total of fifty-four strains of A. calcoaceticus-A. baumannii complex species which were identified to genospecies 2 and 13 by biochemical characteristics was subjected to identify by ribotyping using restriction endonuclease EcoRI, ClaI, and SalI. Of fifty-four strains, twenty-five strains were identified as A. baumannii (genospecies 2) and twenty-one strains as genospecies 13, and six strains changed to genospecies 3, and the rest two strains were confirmed as A. haemolyticus (genospecies 4). This result suggests that the ribotyping may be of value for identification of genospecies and epidemiological information of Acinetobacter strains.
Acinetobacter baumannii* ; Acinetobacter calcoaceticus* ; Acinetobacter* ; Cross Infection ; DNA Restriction Enzymes ; Humans ; Korea ; Molecular Typing ; Ribotyping*