1.Advanced techniques of southern blot hybridization.
In Jang CHOI ; Yong Wook JUNG ; Dae Kwang KIM ; Sung Ik CHANG ; Ihn Hwan LEE
Korean Journal of Anatomy 1991;24(2):219-225
No abstract available.
Blotting, Southern*
2.Diagnosis of human papillomavirus in cervical neoplasia by using southern blot hybridization technique and ViraPap@ HPV DNA detection kit.
Yeon PARK ; Min Soo KIM ; Kee Mook CHUNG ; Jae Hoon CHUNG ; Kyang Hyuk KIM
Korean Journal of Obstetrics and Gynecology 1992;35(10):1501-1508
No abstract available.
Blotting, Southern*
;
Diagnosis*
;
DNA*
;
Humans*
3.Detection of Human Papillomavirus DNA in Condylomata Acuminata Patients using Molecular Hybridization.
Kyoung Chan PARK ; Sang Hak LEE ; Yoo Shin LEE ; Young Kee KIM ; Heung Bae PARK ; Jeong Seon SEO
Korean Journal of Dermatology 1989;27(6):660-665
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
4.The Efficient Transformation of Pleurotus ostreatus using REMI Method.
Joong Ho JOH ; Beom Gi KIM ; Kyo Sun CHU ; Won Sik KONG ; Young Bok YOO ; Chang Soo LEE
Mycobiology 2003;31(1):32-35
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
5.Caveolin-1 is involved in high glucose accelerated human glomerular mesangial cell senescence.
The Korean Journal of Internal Medicine 2017;32(5):883-889
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
6.Detection of herpes simplex virus DNA in cutaneous lesions of erythema multiforme by the polymerase chain reaction.
Korean Journal of Dermatology 1992;30(3):325-333
Herpes simplex virus(HSV) infection has long been considered as a cause of erythema multiform(EM), especially the recurrent type, although its exact role in the pathogenesis of herpes-associated erytherna multiforme(HAEM) is unknown. In this study, HSV DNA was detected in 8 of 29 cases of EM by the polymerase chain reaction(PCR) and in 19 of 29 cases by Southern blot hybridization with PCR products. In 4 cases of EM, we performed PCR & Southern blot hybridization in both lesional and normal skin. HSV DNA was found in 3 of 4 cases of lesional skin but not detected in normal skins by Southern blot hybridization with PCR products. These results confirm the presence of HSV DNA in lesions of HAEM and support the concept of and HSV-specific immune-mediated pathogenesis for this disease.
Blotting, Southern
;
DNA
;
Erythema Multiforme*
;
Erythema*
;
Heme
;
Herpes Simplex*
;
Polymerase Chain Reaction*
;
Simplexvirus*
;
Skin
7.Different Phase of Telomere Shortening with Age in Peripheral Blood Mononuclear Cells.
Ji Seon KANG ; Hoon KOOK ; Won Sang YOON ; Chan Jong KIM ; Tai Ju HWANG
Korean Journal of Pediatric Hematology-Oncology 1999;6(1):31-38
PURPOSE: Telomeres, special protein and tandem repeat DNA structure that cap the ends of linear eukaryotic chromosomes, are essential for chromosome structure and stability. Human telomeric DNA is known to shorten by 30~200 bp with each somatic cell division. However, the phase of telomere changes has not been studied extensively. METHODS: Telomere length was analyzed in the peripheral blood mononuclear cells (PBMCs) of 39 normal controls aged from newborn to 72 years by Southern blot hybridization using PharMingen's TeloQunatTM Telomere Length Assay Kit (Becton Dickinson Co.). RESULTS: The mean telomere length of the population was 9.68 kb (range, 5.65~14.40 kb). The length (kb) decreased with age (A) by the following regression: T=10.86 0.04 A (T=telomere length in kb; A=age in years) (r= 0.38; P=0.016). The mean telomere lengths according to age groups were: 10.26 kb for less than 15 years; 9.92 kb for 16 to 40 years; 8.03 kb for over 40 years. The telomere length of over 40 years was significantly shorter than that of less than 15 years (P=0.013), and than that of 16 to 40 years (P=0.011). The phase of telomere changes was evaluated by age subgroups. The shortening was fastest in individuals of age <5, while the length showed a plateau or slight increment in age group between 5 to 35. The length decreased steadily with age by the regression of 12.43+/-0.07 A (r= 0.500; P=0.034) in age group over 35. CONCLUSION: Telomere length of PBMCs decreases with age, and the different phase of telomere length shortening may suggest that the shortening of telomere is not a constant process over lifespan, but a dynamic process that is differently regulated in age groups.
Blotting, Southern
;
Cell Division
;
Chromosome Structures
;
DNA
;
Humans
;
Infant, Newborn
;
Tandem Repeat Sequences
;
Telomere Shortening*
;
Telomere*
8.Species-Specific Detection of Listeria monocytogenes by Polymerase Chain Reaction.
Byoung Seon YANG ; Jin Sook PARK
Journal of the Korean Society for Microbiology 1997;32(2):213-218
Three Listeria monocytogenes strains isolated from clinical cerebrospinal fluid samples. A PCR was performed by using oligonuclotide primers derived from the invasion-associated protein(iap) related gene sequence. As a result of PCR, we obtained 287-bp DNA fragment of specific product to L. monocytogenes species. The chromsomal DNA of genera Listeria species were subjected to southern blot hybridization with HRP-labeled 287-bp DNA probe to produce a L. monocytogenes banding pattern. In the cases of 10-fold dilution of L. monocytogenes cells, 8 x 10(2) cells/ml were detected by PCR.
Blotting, Southern
;
Cerebrospinal Fluid
;
DNA
;
Listeria monocytogenes*
;
Listeria*
;
Polymerase Chain Reaction*
9.A Case of unusual condyloma Acuminatum in an Immunosuppressed Patient.
Joo Hyun CHOI ; Mi Kyung CHO ; Duk Kyu CHUN ; Kyoung Chan PARK
Korean Journal of Dermatology 1989;27(6):734-737
Condyloma acuminatum is a common viral disease which is transmitted by sexual intercourse. We experienced a case of condyloma acuminatum occurring in a patient of systemic lupus erythematosus who has been treated with various immunosuppressive agents for 10 years. This condylorna acuminatum was unusual in that it was very rapidly growing, recurrent and reluctant to various treatrnents. We investigated the type of human papillomavirus by Southern blot hybridization and schieved partial improvement by bleomycin intralesional injection together with 5-FU topical application.
Bleomycin
;
Blotting, Southern
;
Coitus
;
Fluorouracil
;
Humans
;
Immunosuppressive Agents
;
Injections, Intralesional
;
Lupus Erythematosus, Systemic
;
Virus Diseases
10.Identification of Human Papillomavirus(HPV) in Patients with Cervical Cancer by DNA Hybridization.
Korean Journal of Gynecologic Oncology and Colposcopy 1990;1(1):72-79
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*