No abstract available.
Blotting, Southern*

No abstract available.
Blotting, Southern* ; Diagnosis* ; DNA* ; Humans*

Condylomata acuminata are benign tumors which are mostly venereally transmitted. Common sites were coronal sulcus, perisnal area and prepuce. Among 28 patients, 21 acuminate lesions and 10 papular lesions were found. Twenty eight human genital warts in Korean were analysed by Southern blot hybridization. Sequences related to HPV6/11 are found in 89.3%(25/28) of the condylomata. HPV16 DNA was not found at sll. Subtype of HPV was determined by the restriction pattern of DNA cleaved with PstI restriction enzyme in 7 cases. Six cases of HPV6a and one case of HPV6c are found. The above results suggest that most of condylomata acuminata are caused by HPV6 and HPV11 in Korea.
Blotting, Southern ; Condylomata Acuminata* ; DNA* ; Humans* ; Korea

Restriction enzyme-mediated integration (REMI) was used to transform uracil auxotrophs of Pleurotus ostreatus to prototrophy. When protoplasts of Pleurotus ostreatus were treated by the reaction mixture containing 10 units of BamHI, the frequency of REMI was about 64 transformants per 1 microg of DNA. This efficiency was increased by 14.2 times compared with that of the conventional PEG transformation. The optimal condition for REMI of P. ostreatus was achieved when 1 microg of linearized pTRura3-2 DNA was added into 1x10(7) protoplasts along with 10 units BamHI. Southern blot analysis revealed that about 50% of transformants examined were caused by REMI event and 30% carried single copy insertion at the genome. This suggested that the REMI method might be a useful tool for efficient transformation and tagging mutagenesis of P. ostreatus.
Blotting, Southern ; DNA ; Genome ; Mutagenesis ; Pleurotus* ; Protoplasts ; Uracil

BACKGROUND/AIMS:: We demonstrated the role of caveolin-1 involved in high glucose (HG)-induced glomerular mesangial cells (GMCs) senescence. METHODS:: HG was used to stimulate GMCs. The telomere lengths were analyzed by Southern blot. β-Galactosidase staining was determined. The expressions of caveolin-1 and P53 proteins were determined by Western blot. RESULTS:: Treatment with high concentrations of glucose induced GMC senescence accompanied by shortened telomere length and increase of β-galactosidase staining as well as P53 protein, which was abrogated after application of caveolin-1-siRNA. CONCLUSIONS:: This study proved that HG induced cell senescence in GMCs. The caveolin-1 is involved in HG-induced mesangial cell senescence, and blocking caveolin-1 significantly reduced cell senescence. The effect of caveolin-1 is mediated by P53 pathway.
Aging* ; Blotting, Southern ; Blotting, Western ; Caveolin 1* ; Cell Aging ; Glucose* ; Humans* ; Mesangial Cells* ; Telomere

The PCR primer set JW21-JW22 of Weiss et al. (19), which was reported to amplify a 139-bp fragment of the 16S rRNA gene of Helicobacter pylori, has been recently used for the detection of H. pylori in clinical specimens. However, when we applied JW21-JW22 PCR to other members of the genus Helicobacter and unrelated microorganisms, all of these bacteria produced a 139-bp PCR product. Therefore, we designed a novel primer set, HPU185-HPL826, which produced a 642-bp amplicon of the 16S rRNA gene of H. pylori. Then we further examined the specificity of the novel PCR assay using Southern blot hybridization with an internal probe, HPP225. The PCR assay described in this study was shown to be highly sensitive and specific only to the H. pylori 16S rRNA gene sequences.
Bacteria ; Blotting, Southern ; Genes, rRNA* ; Helicobacter pylori* ; Helicobacter* ; Polymerase Chain Reaction* ; Sensitivity and Specificity

BACKGROUND: The oral ulcer is a common oral disorder, but the precise etiology remains elusive despite of intensive clinical, immunological, hematological and microbiological investigations. OBJECTIVE: The purpose of this study was to examine oral ulcers for the detection of HSV DNA by using PCR and to characterize clinical features of HSV DNA positive cases. METHODS: Specimens collected with cotton swabs and saliva from 48 cases of oral ulcers were examined for HSV DNA by PCR and Southern blot hybridization. RESULTS: 1. HSV DNA was detected in 8 of 48(16.7%) cotton swabbed specimens of oral ulcers and saliva by PCR and Southern blot hybridization. 2. Clinical features of HSV DNA positive oral ulcers were a. predominently located in buccal mucosa. b. mall sized(<3mm), multiple(> or =5) lesions. 3. In Behcet's disease, 2 of 7(28.6%) cases of oral ulcers were positive for HSV DNA CONCLUSION: PCR is an useful and accurate method for the detection of HSV DNA from cotton swabbed specimens of oral ulcers.
Blotting, Southern ; DNA ; Herpes Simplex* ; Mouth Mucosa ; Oral Ulcer* ; Polymerase Chain Reaction* ; Saliva ; Simplexvirus*

Serovars of 22 leptospiral field isolates from rats trapped in Korea were identified by cross-agglutinin absorption test (CAAT). Genomic characteristics of 7 selected isolates and 6 antigenically closely related reference serovars of lai, yeonchon, birkini, gem, mwogolo, and canicola were differentiated by arbitrarily primed PCR (AP-PCR) and southern blot hybridization using 16S rRNA gene probe from Borrelia burgdorferi. Among the 22 isolates, 21 strains were identified as serovar lai by CAAT, while the serological reactivity of NR13 did not accord with that of serovar lai. Results of AP-PCR using primers RSP, KF and PB-1 were in general agreement with those obtained by serological identification, and all 7 isolates including NR13 showed the same profile with serovar lai or yeonchon. In the southern blot hybridization with 16S rRNA gene probe, the isolates were divided into two ribotype groups when HindIII and BamHI digests were employed: isolates NR4, NR13, and serovar lai showed the same profile, and isolates JR34, JR57, KR48, JR77, and JR82 were classified as the another ribotype group. Isolate NR13 and serovar yeonchon, which were isolated in Korea and showed serological differences with serovar lai, were indistinguishable from serovar lai in this DNA study using AP-PCR and ribotyping. These results demonstrate that Korean leptospiral isolates were closely related in DNA level, and ribotyping would be useful for subgrouping of field isolates.
Absorption ; Animals ; Blotting, Southern ; Borrelia burgdorferi ; DNA ; Genes, rRNA ; Korea ; Leptospira* ; Polymerase Chain Reaction* ; Rats ; Ribotyping*

To elucidate the role of plasmid-mediated AmpC-type B-lactamases in clinical practice, cefoxitin-resistant isolates of E. coli (19 strains) and K. pneumoniae (7 strains) from three hospitals in Korea were studied. All of the 26 isolates produced at least one j3-lactamase and 16 (62%) isolates produced AmpC-type B-lactamases poorly inhibited by clavulanic acid. In 16 such isolates, 4 kinds of AmpC enzymes were detected; the pI 8.0 AmpC enzyme in 11 isolates, the pI 8.9 in 3 isolates of E. coli, the pI 8.5 in 1 isolate of E. coli, and the pI 7.8 in 1 isolate of K pneumoniae. The pI 8.0 and 7.8 AmpC enzymes had an apparent molecular mass of 38 kDa and the pI 8.5 and 8.9 AmpC enzymes had a molecular mass of 35 kDa. Cefoxitin resistance was transmissible in six E. coli and three K pneumoniae strains due to a common AmpC-type B-lactamase with a pl of 8.0. This enzyme was confirmed to be CMY-1 B-lactamase by Southern blotting and PCR analysis. Four E. coli isolates produced large amounts of AmpC-type j3-1actamase. They were chromosomal AmpC hyperproducers carrying some alterations in the promoter and attenuator regions of the ampC chromosomal gene. The pI 7.8 AmpC enzyme is currently under study. In conclusion, this study showed that the CMY-1 plasmid-mediated cephamycinase play an important role in cephamycin resistance of K. pneumoniae and E. coli clinical isolates in Korea.
beta-Lactamases* ; Blotting, Southern ; Cefoxitin ; Clavulanic Acid ; Korea ; Pneumonia* ; Polymerase Chain Reaction

BamHI restriction patters and genomic library of Herpes simplex virus type 2 (HSV-2) stram G were constructed, and locations of the glycoproteins gB and gD, and it genes on the fragments were detected by Southern blot analysis. HISV-2 genomic DNAs were cleaved into twenty-seven fragments by BamHI enzyme in the range of 0.72 to 15.08 (total 150.44 kb), which were cloned into the BamHI site of pBluescript SK(+) to construct genome library of the HSV-2. The library was named by the order of the fragment size from smallest one to largest one. HSV-2 glycoprotein gD gene was located in PHLA2-21 and PHLA2-22 recombinant plasmids, gB gene in PHLA2-24 plasmic, and it gene in PHLA2-11 clone by Southern blot analysis.
Blotting, Southern ; Clone Cells ; DNA ; Genomic Library ; Glycoproteins* ; Herpes Simplex* ; Herpesvirus 2, Human* ; Phosphotransferases* ; Plasmids ; Simplexvirus*