• Title/Summary/Keyword: PCR polymorphism

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(CA/GT)n Simple Sequence Repeat DNA Polymorphism in Chlamydomonas reinhardtii (녹조류 Chlamydomonas reinhardtii의 (CA/GT)n Simple Sequence Repeat DNA 다형현상)

  • ;;Marvin W. FAWLEY
    • Korean Journal of Plant Tissue Culture
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    • v.24 no.2
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    • pp.113-117
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    • 1997
  • Simple sequence repeats (SSR) are widely dispersed throughout eukaryotic genomes, highly polymorphic, and easily typed using polymerase chain reaction (PCR). The objective of this study was to determine the polymorphism of different Chlamydomonas reinhartdtii strains and to determine the mode of inheritance of the SSR locus in Chlamydomonas. A genomic DNA library of C. reinhardtii was constructed and screened with a radiolabeled $(AC)_{11}$ probe for the selection of (CA/GT)n repeat clone. Selected clone was seqeuenced, and PCR primer set flanking (CA/GT)n sequence was constructed. PCR was used to specifically amplify the SSR locus from multiple isolates of C. reinhardtii. The locus was polymorphic in some of the C. reinhardtii isolates. However, the locus was amplified only 4 of 6 isolates of C. reinhardtii, not in other 2 isolates of C. reinhardtii, suggesting that this locus is not extensively conserved. A simple Mendelian inheritance pattern was found, which showed 2:2 segregation in the tetrads resulting from a cross between C. reinhardtii and C. smithii. Our results suggest that this simple sequence repeat DNA polymorphism will be useful for identity testing, population studies, linkage analysis, and genome mapping in Chlamydomonas.

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Analysis of Genetic Relatedness in Alternaria species Producing Host Specific Toxins by PCR Polymorphism

  • Kang, Hee-Wan;Lee, Byung-Ryun;Yu, Seung-Hun
    • The Plant Pathology Journal
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    • v.19 no.5
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    • pp.221-226
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    • 2003
  • Twenty universal rice primers (URPs) were used to detect PCR polymorphisms in 25 isolates of six different Alternaria species producing host specific toxins (HST). Eight URPs could be used to reveal PCR polymorphisms of Alternaria isolates at the intra- and inter-species levels. Specific URP-PCR polymorphic bands that are different from those of the other Alternaria spp. were observed on A. gaisen and A. longipes isolates. Unweighted pair-group method with arithmetic mean (UPGMA) cluster analysis using 94 URP polymorphic bands revealed three clustered groups (A. gaisen group, A. mati complex group, and A. logipes group).

Population Analysis of Korean and Japanese Toxic Alexandrium catenella Using PCR Targeting the Area Downstream of the Chloroplast PsbA Gene

  • Kim Choong-Jae;Kim Chang-Hoon;Sako Yoshihiko
    • Fisheries and Aquatic Sciences
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    • v.7 no.3
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    • pp.130-135
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    • 2004
  • The marine dinoflagellate genus Alexandrium, which produces PSP toxins, has a global distribution. As human-assisted dispersal of the species has been suggested, it is important to develop molecular tools to trace the dispersal pathway. To screen population-specific DNA sequences that differentiate Korean and Japanese A. catenella, we targeted the area downstream of the chloroplast psbA gene using PCR with population-specific DNA primers followed by RFLP (restriction fragment length polymorphism) analysis and sequencing. The RFLP patterns of the PCR products divided Korean and Japanese A. catenella regional isolates into three types: Korean, Japanese, and type CMC3, isolated from Korea. We sequenced the PCR products, but found no similar gene in a homology search. The molecular phylogeny inferred from the sequences separated the Korean and Japanese A. catenella strains, as did the RFLP patterns. However, the Japanese isolates included two slightly different sequences (types J and K), while the Korean sequence was the same as the Japanese K type. In addition, a unique sequence was found in the Korean strains CMC2 and CMC3. Population-specific PCR amplification with Japanese A. catenella type-specific PCR primers designed from the type J sequence yielded PCR products for Japanese strains only, showing that the unknown gene can be used for a population analysis of Korean and Japanese A. catenella.

Polymorphism of Insulin-like Growth Factor 1 Receptor Gene in 12 Pig Breeds and Its Relationship with Pig Performance Traits

  • Wang, Wenjun;Ouyang, Kehui;Su, Xifan;Xu, Mingsheng;Shangguan, Xinchen
    • Asian-Australasian Journal of Animal Sciences
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    • v.19 no.11
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    • pp.1541-1545
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    • 2006
  • The polymorphism of insulin-like growth factor I receptor (IGFIR) gene in 12 pig breeds (total n = 593) was detected by PCR-SacII-restriction fragment length polymorphism and allele A (379 bp) or allele B (235 bp and 144 bp) observed. In the studied breeds, it was found that European pigs principally carried allele A, while Chinese native pig breeds principally carried allele B. In addition, the role of pig IGFIR was investigated in 156 Wanbai pigs and 212 Large Yorkshire pigs. Growth related variables including body weight at birth, 2-, 4- and 6-mo of age and backfat thickness and lean percentage estimated by ultrasonography at 6-mo of age were recorded in analyzing the association between IGFIR gene polymorphism and growth traits. AA-genotype pigs exhibited greater (p<0.05) body weights (BW) at birth, 2- and 6-mo of age, but not at 4-mo of age, than those of the BB-genotype in Wanbai and Yorkshire breeds. Moreover, in the Yorkshire breed, AA-genotype pigs had less backfat thickness (p<0.05) and greater lean percentage (p<0.01) than the BB genotype. Based on these results, it is necessary to do more studies on IGFIR before introducing the IGFIR locus into breeding programs.

The Clinical Study on Angiotensin Converting Enzyme Gene Polymorphism in Korean Facial nerve palsy Patients (한국인(韓國人) 구안와사(口眼喎斜) 환자(患者)의 Angiotensin Converting Enzyme 유전자(遺傳子) 다형성(多形成)에 관(關)한 임상연구(臨床硏究))

  • Hong, Jang-mu;Park, Dong-suk;Koh, Hyung-kyun
    • Journal of Acupuncture Research
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    • v.21 no.2
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    • pp.103-113
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    • 2004
  • Objective : This study was designed to investigate the relation between the angiotensin converting enzyme(ACE) gene polymorphism and Facial nerve palsy in the Korean population. Methods : This sudy was carried out on 117 Facial nerve palsy patients who were treated in the department of acupuncture & moxibustion, Hospital of Oriental medical college, Kyung-Hee University and 135 healthy control subjects. Blood samples from all subjects were obtaind for DNA extraction. The extracted DNA was amplified by polymerase chain reaction(PCR). PCR products were visualized by 2% agarose gel electrophoresis. Results : The sub-genotypes of ACE gene were II homozygotes, ID heterozygotes, DD homozygotes. While the distribution of ACE polymorphism in control subjects was 33%, 43%, 24%, the distribution of it in Facial nerve palsy patients was 32%, 50%, 18%(II, ID, DD). Thus, there was no significant different between the control and Facial nerve palsy groups. Conclusions : we conclude that there is no significant association between ACE gene polymorphism and Facial nerve palsy in Korean population. However, the findings of this study need to be confirmed in more patients and further studies. Additional epidemiologically based studies of the effects and relationship between ACE or other genes and lifestyles with regard to Facial nerve palsy is required.

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Frequency of the Angiotensin - Converting Enzyme (ACE) Gene Polymorphism in the General Population and the Elite Endurance Students in Korea

  • Choung, Ho-Jin;Yoon, Song-Ro;Choi, Soo-Kyung
    • Journal of Genetic Medicine
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    • v.3 no.1
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    • pp.11-13
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    • 1999
  • Recently it was reported that Insertion/Deletion polymorphism in the gene coding for Angiotensin-Converting Enzyme (ACE) is associated with human capacity for physical performance. This study was performed to genotyping of the ACE gene to determine the correlation between elite endurance performance and ACE I/D gene polymorphism. DNA sample was obtained from peripheral blood, hair roots and mouth epithelial cell in 739 general population and 200 elite athletic performance students. The ACE gene was amplified by polymerase chain reaction (PCR) using allele specific oligonucleotide primers. 155, 525 bp and 237 bp PCR products indicating the presence of insertion(I) and deletion(D) alleles, respectively, were clearly resolved after electrophoresis on a 2% agarose gel with ethidium bromide. Of the 200 elite athletic performance population subjects, 68(34%) showed ACE genotype 11,100(50%) genotype ID and 32(16%) genotype DD. Of the 739 general population subjects, 259(35.1%) showed ACE genotype 11,363(49.1%) genotype ID and 117(15.8%) genotype DD. Therefore ACE I/D gene polymorphism was not associated with human capacity for physical performance.(p>0.05)

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No association of Polymorphism in the Estrogen-synthesizing Enzyme Genes CYP19 with Advanced Endometriosis in Korean Women (한국인 여성에서 중증 자궁내막증의 발생 위험도와 CYP19 유전자 다형성과의 관련성에 관한 연구)

  • Hur, Sung Eun;Lee, Ji Young;Moon, Hye-Sung;Chung, Hye Won
    • Clinical and Experimental Reproductive Medicine
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    • v.32 no.2
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    • pp.171-176
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    • 2005
  • Objective: To investigate whether polymorphism of CYP19 gene is associated with the risk of advanced endometriosis in Korean women. Methods: Blood samples were collected from 202 endometriosis patients and 221 controls. The patients with endometriosis of stages III and IV diagnosed by both pathologic and laparoscopic findings to according modified AFS classification. The women undergoing laparoscopic surgery or laparotomy for non-malignant lesions were included in the control group. Polymerase chain reaction (PCR) and restriction fragment length polymorphism (RFLP) of PCR products were done to determine all individuals' genotype. Results: The heterozygous allele in CYP19 gene was the most common genotypes in both endometriosis and healthy control groups (52.0% vs. 46.1%). CYP19 gene polymorphisms did not show the significant differences between the control group and endometriosis group. Conclusion: The results suggested that the CYP19 genetic polymorphism was not associated with a risk of advanced endometriosis in Korean women.

Detection of Single Nucleotide Polymorphism in Human IL-4 Receptor by PCR Amplification of Specific Alleles

  • Hwang, Sue Yun;Kim, Seung Hoon;Hwang, Sung Hee;Cho, Chul Soo;Kim, Ho Youn
    • Animal cells and systems
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    • v.5 no.2
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    • pp.153-156
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    • 2001
  • A key aspect of genomic research in the “post-genome era”is to associate sequence variations with heritable phenotypes. The most common variations in the human genome are single nucleotide polymorphisms (SNPs) that occur approximately once in every 500 to 1,000 bases. Although analyzing the phenotypic outcome of these SNPs is crucial to facilitate large-scale association studies of genetic diseases, detection of SNPs from an extended number of human DNA samples is often difficult, labor-intensive and time-consuming. Recent development in SNP detection methods using DNA microarrays and mass spectrophotometry has allowed automated high throughput analyses, but such equipments are not accessible to many scientists. In this study, we demonstrate that a simple PCR-based method using primers with a mismatched base at the 3'-end provides a fast and easy tool to identify known SNPs from human genomic DNA in a regular molecular biology laboratory. Results from this PCR amplification of specific alleles (PASA) analysis efficiently and accurately typed the Q576R polymorphism of human IL4 receptor from the genomic DNAs of 29 Koreans, including 9 samples whose genotype could not be discerned by the conventiona1 PCR-SSCP (single strand conformation polymorphism) method. Given the increasing attention to disease-associated polymorphisms in genomic research, this alternative technique will be very useful to identify SNPs in large-scale population studies.

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Detection of Mycobacterium avium ssp paratuberculosis in Korean Cattle by the Polymerase Chain Reaction (한우 혈액에서 PCR을 이용한 Mycobacterium avium ssp paratuberculosis의 검출)

  • Kim, Kwang-Hyun;Kwak, Kil-Han;Song, Hee-Jong;Cho, Jeong-Gon
    • Journal of Veterinary Clinics
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    • v.27 no.1
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    • pp.23-28
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    • 2010
  • Mycobacterium avium ssp paratuberculosis, intracellular bacteria that can cause chronic granulomatous enteritis in cattle, continues to pose significant economic losses and health problem with high prevalence. The purpose of this study is the polymerase chain reaction (PCR)-base strategy for early detection of M. avium ssp paratuberculosis in whole blood. Blood samples were collected from korean cattles in Jeonbuk, Korea. The 16 out of 88 serum samples were detected M. partuberculosis by ELISA. Then samples of infected 8 Korean cattles were amplified by PCR. The PCR amplified targets are 16s rDNA and heat shock protein 65kDa (hsp 65). The 16s rDNA provided a highly sensitive and specific tool for the direct detection of mycobacteria. In addition M. avium was confirmed characteristically by the hsp65. Finally there were sure to M. avium ssp paratuberculosis by IS900 PCR. The restriction fragment length polymorphism was identified by PCR amplifications and subsequence restriction enzyme digestions with Pst I of a hsp65. These results indicate that confirm M. avium with 16s rDNA, hsp65 and a restriction fragment length polymorphism in the hsp65 gene can be seem the other pattern. Therefore, these results can be used for clinical direct detections of M. avium ssp paratuberculosis in whole blood of Korean cattle and also to be used epidemiological researches.

Association between a p73 Gene Polymorphism and Genetic Susceptibility to Non-small Cell Lung Cancer in the South of China

  • Wang, Shuang-Shuang;Guo, Hai-Yan;Dong, Lin-Li;Zhu, Xiang-Qian;Ma, Liang;Li, Wen;Tang, Jian-Xin
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.23
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    • pp.10387-10391
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    • 2015
  • Background: This study aimed to identify any association between the p73 gene G4C14-to-A4T14 polymorphism and risk of non-small cell lung cancer (NSCLC) in the south of China. Materials and Methods: We genotyped the p73 gene polymorphism of peripheral blood DNA from 168 patients with NSCLC and 195 normal controls using HRM (high resolution melting) and PCR-CTPP (polymerase chain reaction with confronting two-pair primers). Results: The results of genotyping by HRM and PCR-CTPP were consistent with direct sequencing, the p73 genotype distribution in 168 lung cancer patients being as follows: GC/GC 101 cases (60.1%), GC/AT 59 cases (35.1%), AT/AT 8 cases (4.8%). The carriers of AT/AT genotype had a significantly reduced risk of NSCLC (OR=0.370; 95%CI: 0.170-0.806; p=0.010) as compared with non-carriers. However, we found no relations between p73 genotypes and histological type (p=0.798, $x^2=0.452$), tumor stage (p=0.806, $x^2=0.806$), or lymph node metastasis (p=0.578, $x^2=1.098$). Conclusions: Our findings suggest that the p73 G4C14-to-A4T14 polymorphism may be a modifier of NSCLC susceptibility in the Chinese population.