• Parasites, Hosts and Diseases
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• v.39 no.1
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• pp.43-48
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• 2001

We have studied the genetic differences among four isolates of Trichinella including a new strain of Trichinella spiralis (ISS 623) recently found from a human case who took a badger in Korea. Because they have a different host origin and came from geographically separated regions, we supposed the genetic pattern of the isolates might be different as had been previously reported. It was analysed by PCR-RFLP analysis of the rDNA repeat that can readily distinguish a species or strain from others. Isolated genomic DNA of each isolate of Trichinella larvae was amplified with ITSl specific primers and digested with restriction endonucleases. The PCR product of ITSl was confirmed using Southern blot analysis to be a 910 Up fragment. The restriction fragments of each isolate had variable patterns when it was digested with Rsa I only. According to the RFLP patterns, the estimated genetic divergence between each isolate was different. In conclusion, four isolates of Thichinella including a new strain of T. spiralis obtained from a Korean patient may have genetic differences in the ITSl region and the Shanghai isolate was genetically more similar to the Japanese unknown isolate than others in the ITSl region.

• Proceedings of the Korean Society for Food Science of Animal Resources Conference
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• 2004.05a
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• pp.177-180
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• 2004

본 연구는 PCR-RFLP 및 PCR-SSCP 기법을 이용하여 PSE 돈육을 생산하는 PSS 돼지 유전자 진단 기술을 개발하고 이를 이용한 국내 종돈 및 교잡 비육돈의 PSS 유전자 출현 빈도를 파악하고자 수행하였다. 돼지 PSS의 원인이 되는 ryanodine receptor 유전자의 단일염기 돌연변이 $C{\rightarrow}T$ ; $Arg\;{\rightarrow}\;Cys$)를 포함하는 134 bp 영역을 PCR로 증폭한 후 RFLP 및 SSCP 기법으로 분석한 결과 동형접합체의 정상(N/N), 이형접합체의 잠재성 개체 (N/n) 그리고 돌연변이 유전자를 동형접합체 상태로 갖는 PSS 감수성 개체(n/n)에 각각 특이적인 유전자형이 검출되었다. 특히, PCR-SSCP기법을 이용한 RYR1 유전자 돌연변이 검출 방법은 보다 신속 간편하면서도 상대적으로 분석비용이 저렴한 정확성이 높은 PSS 돼지 진단기술로서 대규모 돼지집단검색이나 RFLP 방법으로 판정이 불확실한 시료의 재검에 효율적으로 이용할 수 있을 것으로 판단된다.

• Archives of Pharmacal Research
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• v.23 no.4
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• pp.418-423
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• 2000

Oka strain VR-795 (Varicella Zoster Virus, VZV) of American Type Culture Collection (ATCC) has been used for chickenpox vaccine production. In order to use this strain for vaccine production, the strain must be identified and its stability must be confirmed. The identification of the Oka strain has been confirmed using Restriction Fragment Length Polymorphism (RFLP) and DNA sequence analysis of glycoprotein-II (gp-II). The amino acid sequences of Oka deduced from the DNA sequence of gp-II have changed at three amino acids against Ellen and at one amino acid against Webster. To prove the stability of the Oka strain during the passage, RFLP and DNA sequence analyses were also used with 11, 15 and 23 times of virus passage. We found that the Oka strain was stable at passages of up to 23 times, based on the RFLP and DNA sequence analyses. The confirmed Oka strain was renamed as BR-Oka for the purposes of chickenpox vaccine production.

• Environmental Mutagens and Carcinogens
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• v.22 no.1
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• pp.30-38
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• 2002

In view of the effect of antithrombin III on essential hypertension, we investigated the 5' untranslated exon 1 length (I/D) polymorphism and intron 5'-DdeI RFLP of the human antithrombin III gene in the Korean patients with essential hypertension. There were no significant differences in the allele, genotype and haplotype frequencies of these polymorphisms between normotensive and essential hypertensive subjects. The significant linkage disequilibrium was however, detected between I/D polymorphism and Dde I RFLP. The I/D polymorphism was also significantly associated with BMI, total cholesterol (TC) and HDL- cholesterol levels, while DdeI polymorphism with age and BMI. Therefore, our results suggest that the significant association between the genetic polymorphisms in the antithrombin III gene and variable cardiovascular risk factors may reflect the potential role of human antithrombin III gene in cardiovascular function.

• Animal cells and systems
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• v.4 no.1
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• pp.87-90
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• 2000

Essential hypertension is a multifactorial disease, and has been shown to be associated with insulin resistance. The relationship between the genetic variation of the insulin receptor (INSR) gene and essential hypertension In Korean population was investigated by the Nsi 1 restriction fragment length polymorphism (RFLP) pattern of this gene. The observed genotype frequencies of INSR gene were not deviated from those expected for the Hardy-Weinberg equilibrium (HWE), but a significant association was observed between essential hypertension and N1 allele of Nsi 1 RFLP at the INSR gene ($X^2$-test; P<0.05). Moreover, the frequency of N1 allele was significantly different between normotensives and essential hypertensives in subgroups that were not obese ($X^2$-test; P<0.05). These data suggest that the Nsil RFLP of INSR gene may be a useful genetic marker for essential hypertension in Korean population.

• Tuberculosis and Respiratory Diseases
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• v.76 no.2
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• pp.59-65
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• 2014

Background: Variable-number tandem repeat (VNTR) typing is a promising method to discriminate the Mycobacterium tuberculosis isolates in molecular epidemiology. The purpose of this study is to determine the optimal VNTR combinations for discriminating isolated M. tuberculosis strains in Korea. Methods: A total of 317 clinical isolates collected throughout Korea were genotyped by using the IS6110 restriction fragment length polymorphism (RFLP), and then analysed for the number of VNTR copies from 32 VNTR loci. Results: The results of discriminatory power according to diverse combinations were as follows: 25 clusters in 83 strains were yielded from the internationally standardized 15 VNTR loci (Hunter-Gaston discriminatory index [HGDI], 0.9958), 25 clusters in 65 strains by using IS6110 RFLP (HGDI, 0.9977), 14 clusters in 32 strains in 12 hyper-variable VNTR loci (HGDI, 0.9995), 6 clusters in 13 strains in 32 VNTR loci (HDGI, 0.9998), and 7 clusters in 14 strains of both the 12 hyper-variable VNTR and IS6110 RFLP (HDGI, 0.9999). Conclusion: The combination of 12 hyper-variable VNTR typing can be an effective tool for genotyping Korean M. tuberculosis isolates where the Beijing strains are predominant.

• The Plant Pathology Journal
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• v.38 no.6
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• pp.616-628
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• 2022

Resistance to pyraclostrobin due to a single nucleotide polymorphism at 143rd amino acid position on the cytochrome b gene has been a major source of concern in red pepper field infected by anthracnose in Korea. Therefore, this study investigated the response of 24 isolates of C. acutatum and C. gloeosporioides isolated from anthracnose infected red pepper fruits using agar dilution method and other molecular techniques such as cytochrome b gene sequencing, polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP), and allele-specific polymerase chain reaction (PCR). The result showed that four isolates were resistant to pyraclostrobin on agar dilution method and possessed GCT (alanine) codon at 143rd amino acid position, whereas the sensitive isolates possessed GGT (glycine). Furthermore, this study illustrated the difference in the cytochrome b gene structure of C. acutatum and C. gloeosporioides. The use of cDNA in this study suggested that the primer Cacytb-P2 can amplify the cytochrome b gene of both C. acutatum and C. gloeosporioides despite the presence of various introns in the cytochrome b gene structure of C. gloeosporioides. The use of allele-specific PCR and PCR-RFLP provided clear difference between the resistant and sensitive isolates. The application of molecular technique in the evaluation of the resistance status of anthracnose pathogen in red pepper provided rapid, reliable, and accurate results that can be helpful in the early adoption of fungicide-resistant management strategies for the strobilurins in the field.

• Pediatric Gastroenterology, Hepatology & Nutrition
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• v.7 no.2
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• pp.137-142
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• 2004

Purpose: The resistance of H. pylori to clarithromycin is one of the major causes of eradication failure. In H. pylori, clarithromycin resistance is due to point mutation in 23S rRNA. The aims of this study were to investigate the mutation of 23S rRNA and to examine the association of cagA, vacA genotype and clarithromycin resistant genes. Methods: H. pylori DNA was extracted from antral biopsy specimens from 27 children with H. pylori infection. Specific polymerase chain reaction (PCR) assays were used for cagA and vacA. Mutations associated with clarithromycin resistance were detected by using PCR restriction fragment length polymorphism (RFLP) analysis of 23S rRNA gene. Results: A2143G mutation was detected in one case and A2144G in 4, indicating 18.5% were clarithromycin resistant. Among the total of 27, cagA was present in 25 (93%), vacA s1a/m1 in 6 (22%), s1a/m2 in 3 (11%), s1c/m1 in 16 (59%), and s1c/m2 in 1 (4%). All of the 5 clarithromycin resistant strains were cagA (+), among which 2 were s1a/m1 and 2 were s1c/m1. There was no relation between genotypes and clarithromycin resistant genes. Conclusion: Detection of H. pylori resistance to clarithromycin using PCR RFLP from biopsy specimens might be useful for the selection of antibiotics. Clarithromycin resistant genes are not associated with genotypes of cagA and vacA.

• Journal of Animal Science and Technology
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• v.46 no.3
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• pp.295-300
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• 2004

The identification of the leptin gene in 1994 and it's adipocytes specific protein leptin hal provided the first physiological links to the regulatory system controlling body weight and fat deposits. The meat tastes is mainly determined by quantifY and quality of triglyceride stored in adipose tissue. This study was conducted to analyze genetic cbaracteristics of Hanwoo leptin gene and also to investigate the association of DNA marlcer with some economic meat traits for Hanwoo. The leptin hormone gene polymorphisms were identified by digestion with Kpn2 I and Msp I. Slaughter weight(SWI), slaughter peroentage(SP), longissimus muscle area(LMA), beef marbling score(MS) and back fat thickness(BF) were compared among three genotypes by P(R..RFlJ> and showed significant differences among genotypes. PCR-RFLP(Kpn2 I) were detected significant for SP, MS and BF. The allele was essociated with fatter carcasses and C allele with leaner carcasses.

• The Korean Journal of Mycology
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• v.26 no.2 s.85
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• pp.173-181
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• 1998

To identify genetic difference of 13 strains in three Pleurotus species, analyses of rDNA, AP-PCR and RFLP were carried out. IGRI and $ITSI{\sim}II$ regions of rDNA amplified by PCR were about 0.9 and 0.7 kb, respectively. These PCR products were digested with six restriction enzymes to analyse polymorphism. Especially, treatment of HaeIII enzyme on $ITSI{\sim}II$ regions showed specific bands in three Pleurotus sajor-caju strains. Genetic differences among three species were classified by similarity analyses based on rDNA polymorphism. Various band patterns of $2,500{\sim}150\;bp$ were showed by AP-PCR. Identification of species and varieties in 13 Pleurotus strains was possible according to primers used in AP-PCR. In order to develop genetic markers, RFLPs using IGRI and $ITSI{\sim}II$ probes derived from ASI 2180 and 2070 were carried out on eight Pleurotus varieties. RFLP patterns using IGRI probe were more various than that of $ITSI{\sim}II$ probe.