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

Genomic DNAs were extracted from cervical lavages of 49 patients with cervical cancer. Dot and Southern blot hybridization were performed using the P-labeled HFV DNA probes to find high risk HPV(type 16 and 18) infection that is known as the mast prevalent pathogenic factor in cervical cancer. Furthermore, genornic DNAs purified frnm cervical cancer tissues were studied in 8 out of 49 patients allowing us to convince the results from cervical lavages. The results were as follaws: 1. Dot blot analysis were used to examine the sensitivity and specificity of hybridization condition and HPV-DNA probes. Fasitive signals were obtained even at the level of 10pg for HPV DNA, but no signals could he detected at the level of as much as 400pg for salmon sperm DNA. 2. Dot blot of DNAs from cervircal lavages showed positive signals in 32.7%(16/49) with HPV type 16 probe and 20.4% (10/49)and one mixed infection was found. 3. When the DNAs from cervircal lavages of 49 patients were classified according to the clinical stage of cervical cancer, the infection rates of HPV type 16 and 18 were 50% (2/4) in CIN, 80% (4/5) in stage I, 64. 2% (9/14) in stage I b, 45% (9I20) in stage II and 16. 7% (1/6) in stage Ill and K respectively. The occurrenr,e of HPV type 16 and 18 seemed to be the highest in the cervical cancer stage 1 (68.4%(13/19). 4. Experiments perfornecl with genomic DNAs from 8 cancer tissues showed similar results compared to those of cervical lavages, but the intensity of positive signals was stronger. 5. Genomic DNAs from 5 patients(3 cases from cervical lavages and 2 cases from cervical cancer tissues) which showed strong positive signals to the dot blot analysis were further examined by Southern blot hybridixation using HFV type 16 DNA probe. When DNAs were digested with Pst 1 restriction enzyme, the five characteristic frgmenta of BFV type 16(2.8, l.9, l.6, 1.0 and 0.5 kb long in length) were recognized in ell 5 cases, These results may suggest a direet relatianship between HPV type 16 & 18 infectioas considered as the most effective methods for HPV detectioe and typing. Mo1ecular biclogieal studies in the reserarch of HPV are expected to reveal and help us understand the pathogenesis of cervical cancer.
Blotting, Southern ; Coinfection ; DNA Probes ; DNA* ; Humans* ; Salmon ; Sensitivity and Specificity ; Spermatozoa ; Therapeutic Irrigation ; Uterine Cervical Neoplasms*

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*

The purpose of this study is to develop species-specific DNA probe for detection and identification of Prevotella intermedia (P. intermedia) G8-9K-3. This study procedure includes (1) whole-genomic DNA extraction of P. intermedia G8-9K-3 (2) construction of the genomic DNA library, (3) screening of strain-specific DNA probe by reverse dot hybridization, (4) confirmation of strain-specific DNA probe by Southern blot hybridization, (5) determination of nucleotide sequences of strain-specific DNA probe. Twenty-eight recombinant plasmids containing Hind III-digested DNA fragments of P. intermedia G8-9K-3 were obtained. Reverse dot Hybridization and Southern blot analysis data showed that one of them, Pig3, could be P. intermedia G8-9K-3-specific DNA probe. This datum indicates that this Pig3 DNA probe could be useful in detection and identification of the P. intermedia G8-9K-3 strain.
Base Sequence ; Blotting, Southern ; DNA* ; Gene Library ; Mass Screening ; Plasmids ; Prevotella intermedia* ; Prevotella*

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*

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*