• Journal of Ginseng Research
• /
• 제14권2호
• /
• pp.310-316
• /
• 1990

This study was focused on the characterization of mitochondrial DNA (mtDNA) for molecular 9enetical approach of energy Production related mechanism in Panax ginseng. The simple and efficient method of mtDNA isolation from ginseng has been developed by modification of recently advanced methods. This procedure can successfully apply to mtDNA isolation of several plants. mtDNA of etiolated shoot and one-year root were digested with restriction endonucleases, but that of 6-year root not. Any difference was not observed in the restriction endonuclease digestion patterns among the ginseng variants. Molecular size of ginseng mtDNA was estimated at least 159 kb by the restriction endonuclease fragment analysis. The 4.5 kb extra band at the lane of EcoRII treatment could be observed in restriction patterns digested with the methylation sensitive endonucleases, BstN I and EcoRII. For construction of mitochondrial genomic library of ginseng, mtDNA was partially digested with EcoRl, and packaged with EMBL4 phage vector. Genomic library was screened and purified for further research including restriction mapping of ginseng mtDNA, and cloning of the genes. The gene of ATP synthase A subunit was cloned from the purified EMBL4 library clone No. 16. Now, clone No. 16 is subcloned for structure gene sequence analysis.

• 한국수산과학회지
• /
• 제30권5호
• /
• pp.804-808
• /
• 1997

한국산 참굴의 유전적 특성을 조사하기 위하여 한국의 지역별 참굴을 대상으로 mtDNA 제한효소 절편분석과 클로닝을 수행하였다. 지역별로 각각 20개체의 mtDNA에 대하여 8가지 제한효소를 사용하여 DNA 절편양상을 분석한 결과 서해안산 참굴에서는 개체간 차이가 없는 단일 양상이었으며 남해안산의 경우는 두 가지 양상을 보였으며 차이는 HindIII 절단양상에서만 나타났다. 그 중 소수 개체들에서 나타난 양상은 서해안산 개체들에서의 양상과 동일하여 남해안에 서해안산 참굴이 유입되어 혼재하는 것으로 추정되었다. 한국산 참굴의 mtDNA를 대장균 E. coli HB101에서 클로닝하여 유전적 분석을 용이하도록 하였다. 전체 mtDNA를 제한효소를 사용하여 세부분으로 나누어 pUC19 유전자운반체에 클로닝하였다. 클로닝된 재조합 DNA를 제한효소들로 절단하여 한국산 참굴 mtDNA의 제한효소지도를 작성하였다. 남해안산과 서해안산 참굴 mtDNA에서 HindIII 적단 양상이 다르게 나타남을 확인하였고 이는 남해안산이나 서해안산에서 염기치환의 돌연변이에 의한 것으로 사료되었다.

• 대한미생물학회지
• /
• 제22권4호
• /
• pp.463-471
• /
• 1987

The genomes of leptospiral field isolates from Korea belonging to serogroup Icterohaemorrhagiae (21 strains) and serogroup Canicola (1 strain) were analysed and compared by restriction enzyme analysis with EcoRI and HindIII as digesting enzymes. One isolate belonging to serogroup Canicola showed the same pattern as serovar portlandvere. All 21 isolates belonging to serogroup Icterohaemorrhagiae showed almost same patterns as Leptospira serovar lai from China, But with very slight differences 21 isolates could be classified into 8 subtypes and these grouping seems to reflect the differences in epidemiological niche. And also the geographical data consisted with the grouping into 8 subtypes. According to our results, we concluded that the restriction endonuclease analysis of chromosomal DNA will be an accurate and reliable method to compare and classify pathogenic leptospires.

• 고려인삼학회:학술대회논문집
• /
• 고려인삼학회 1990년도 Proceedings of International Symposium on Korean Ginseng, 1990, Seoul, Korea
• /
• pp.168-174
• /
• 1990

This study was focused on the characterization of mitochondrial DNA (mtDNA) for molecular genetically approach of energy Production related mechanism in Panax Ein.fend. The simple and efficient method of mtDNA isolation from ginseng has been developed by modification of recently advanced methods. This procedure can successfully apply to mtDNA isolation of several plants. MtDNA of etiolated shoot and one-year root were digested with restriction endonucleases, but that of 6-year root not Any difference was not observed in the restriction endonuclease digestion patterns among the ginseng variants. Molecular size of ginseng mtDNA was estimated at least 159 kb by the restriction endonuclease fragment analysis. The 4.5 kb extra band at the lane of EcoRll treatment could be observed in restriction patterns digested with the methylation sensitive endonucleases, BstN 1 and EcoRll. For construction of mitochondrial genomic library of ginseng, mtDNA was partially digested with EcoRl, and packaged with EMBL4 phage vector Genomic library was screened and purified for further research including restricttion mapping of ginseng mtDNA, and cloning of the genes. The gene of ATP synthase A subunit was cloned koto the purified EMBL4 library clone No. 16. Now, clone No. 16 is subcloned for structure gene sequence analysis.

• Restorative Dentistry and Endodontics
• /
• 제20권2호
• /
• pp.577-609
• /
• 1995

Bacteria have been regarded as one of the most important factors in pulpal and periapical diseases. Streptococci are frequently isolated facultative anaerobes in infected root canals. Recently molecular biological techniques have been rapidly progressed. This study was designed to apply the molecular biological tools to the identification and classification of streptococci in the endodontic microbiology. Streptococci isolated from infected root canals were identified with both Vitek Systems and API 20 STREP. Identification results were somewhat different in several strains of streptococci. Eighteen streptococci and enterococcal was difficult so to digest plasmid DNA using Hind III and EcoRI to differentiate strains by restriction enzyme analysis of plasmid DNA. 16S rDNA of chromosome was amplified by polymerase chain reaction(PCR) and then restricition fragment length polymorphism(RFLP) using several restriction enzymes was observed. The molecular mass of 16S rDNA of chromosomal DNA was approximately 1.4kb. There were three to five RFLP patterns using eight restriction enzymes. RFLP patterns digested with CfoI which recognizes four base sequences were identical in all stains. Hind III which recognizes six base sequences could not digest the 16S rDNA. Restriction enzymes which recognize five base sequences were suitable for RFLP pattern analysis. At least three different restriction enzymes were needed to compare each strains. 16S rDNA PCR-RFLP was simple and rapid to differentiate and classify strains and could be used in the epidemiological study of root canal infections.

• 환경생물
• /
• 제18권1호
• /
• pp.53-61
• /
• 2000

서울시 소재 지하수 중 중금속으로 오염되어 음용수 이외의 생활 용수로 사용하고 있는 1개 정점과 음용수로 사용하고 있는 1개 정점, 대조군으로 강화도 천연 동굴의 1개 정점을 대상으로 실험을 실시하였다. 지하수 세균군집의 유전적 다양성의 변화를 보기 위해 지하수 세균 군집에서 16SrDNA를 증폭하는 primer로 PCR(polymerase chain reaction)을 실시한 후 ARDRA(amplified ribosomal DNA restriction analysis) 지문 분석으로 비교하였다. 16S rDNA를 증폭하여 제한효소 지문분석을 한 결과 in situ와 음용수에서 유전적 다양성이 상대적으로 크게 나타났다. 지하수 세균 군집의 ARDRA지문 분석은 상이한 환경과 서식지를 반영한 유전적 차이를 빠르게 비교 분석할 수 있었으며 지하수의 오염도에 따른 미생물의 다양성(천연 동굴>음용수>오염수)을 확인할 수 있었다.

• 대한수의학회지
• /
• 제33권1호
• /
• pp.43-51
• /
• 1993

Mitochondrial DNA(mt DNA) of Mammalian is the circular one which the 16.5K base pairs and show the maternal inheritance. Evolutional speed of nucleotide sequence is very fast. So that polymorphic analysis of mt DNA provide the useful informations to investigate the genetic relations of interspecies. Authors trials were focussed to compare with the polymorphic differences of mitochondrial DNA between Jindo and Japanese mongrel dogs. DNA was extracted from bloods of 21 head of Jindo dogs and 20 head of Japanese dogs and isolated using 10 kinds of restriction endonucleases(Apa I, BamH I, Bgl II, EcoR I, EcoR V, Hinc II. Hind III, Pst I, Sty I, Xba I) and then separated by the agarose gel electrophoresis. After sourthern blotting hybridization was completed using the mtDNA of Japanese mongrel dogs as a probe. Autoradiography was used to compare the polymorphism of mtDNA both dogs. The results obtained were as follows; 1. mt DNA of Jindo dog showed polymorphism resulting cleavage with four kinds of restriction endonuclease, Apa I, EcoR V, Hinc II, Sty I. While in the Japanese mongrel dogs observed the polymorphism in the five kinds of restriction endonuclease supplemented with EcoR I. 2. Compared with both dogs the frequency differences of DNA polymorphism were recognized in the specific restriction endonuclease Apa I. Consequently in the restriction endonuclease Apa I both dogs classified with three types as A, B, C however in the Jindo dogs frequency of C type was 71.5 percent but in Japanese mongrel dogs observed 45 percent in the A type. 3. DNA polymorphism obtained from the use of five kinds of restriction endonuclease were classified with seven types. In Jindo dogs frequency was highest in the type 6 as 71.4 percent but in the Japanese mongrel dogs showed 35 percent in the type 5. 4. Genetic distances calculated by NEI method showed 0.0089 in Jindo dogs and was 0.0094 in the Japanese mongrel dogs.

• 미생물학회지
• /
• 제23권2호
• /
• pp.115-121
• /
• 1985

Lactobacillus casei에 감염하는 독성 phage중 각 분류꾼을 대표하는 5종의 phage (J1, TK93, K1, PD 5 빛 CP 1)와 l 종의 용원 phage (${\phi}$ 1043) 의 핵산의 특성을 바교 검토하였다. 실험한 6 종의 phage는 모두 double S stranded DNA를 갖고 있였으며 J1. TK93, K1 및 ${\phi}$ 1043 DNA의 크기는 약 42Kb, PD5 와 CP1 DNA는 140K Kb정도로 서로 비슷하였다. EcoR 1으로 절단시 J1, TK93, K1, PD5, CP1 및 ${\phi}$1043은 각각 13, 13. 11, 14, 14와 12개의 절편을 갖는 특징적인 절단양식을 보여주었다. J1, TK93 및 ${\phi}$ 1043 DNA에는 cohesive end가 존재 하였고 K1, PD5 빛 CP1 DNA에는 없는 것으로 사료되었다. J1과 TK93 DNA의 제한효소 지도를 작성하여 비교 검토하였으며 이상의 결과로부터 진화적 유연관계를 검토하였다.

• Animal cells and systems
• /
• 제3권1호
• /
• pp.73-78
• /
• 1999

Although there are several methods for the preparation of mitochondrial DNA (mtDNA) from mammalian tissues, most are relatively long ultracentrifugation or manipulations by a small-scale method. We escribed a rapid method for large-scale extraction of mtDNA from human placental and horse liver tissues. The method is based on the preparation and homogenization of tissues, urification of crude mitochondria by differential centrifugations and isolation of mtDNA by alkaline Iysis. It was improved from Pre-existing methods by replacing some steps with simpler ones and discarding many others. This method gives a high yield of pure mtDNA(approximately 1-5mg from one placenta; ca. 400-600 g wet weight), depending on its sources (fresh tissue gave better results than frozen one). The resulting mtDNA indicated that this method can yield mtDNA in sufficient purity and quantity to identify the direct restriction analysis on agarose gel, random-primed labeling as a probe, and end labeling. Therefore, the method is ideal for obtaining good mtDNA samples to conduct routine restriction fragment length polymorphism (RFLP) analyses of natural populations for genetic studies.

• 한국수산과학회지
• /
• 제27권5호
• /
• pp.613-619
• /
• 1994

황해에 서식하는 참조기(Pseudosciaena polyactis Bleeker) 각 계군의 유전적 차이점을 분석하기 위하여 중국에서 3지역(Zhoushan, Shanghai, Qingdao), 한국 2지역(목포, 인천)에서 채집된 참조기로부터 mitochondrial DNA(mtDNA)의 RFLP(제한효소 절편 다형현상)를 분석하였다. 총 18종의 제한효소를 이용하여 처리한 결과 5개 집단 모두 동일한 크기인 $16.9{\pm}0.6kb$의 mtDNA를 소유한 것으로 나타났으며 이는 다른 어류군들과 유사한 크기였다. 참조기 mtDNA에 대한 RFLP 분석을 행한 결과 각 집단 마다 대략 40여개의 절편이 관찰되었고 5개 집단 모두 동일한 mtDNA 절편양상을 보였으나 사용된 제한효소 중 좌ApaI, EcoRI, PstI, SstII 및 SmalI에서 중국과 한국 집단내 또는 집단간 절편 양상의 차이도 관찰할 수 있었다.