• Title/Summary/Keyword: PCR/RFLP analysis

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Candidatus Phytoplasma trifolii Associated with Witches' broom of Lespedeza cyrtobotrya M.

  • Kim, Young-Hwan;Jung, Hee-Young
    • The Plant Pathology Journal
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    • v.23 no.2
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    • pp.106-108
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    • 2007
  • The Symptoms of witches' broom disease caused by phytoplasma including general stunting and yellowing, were observed in leafy lespedeza (Lespedeza cyrtobotrya M.) on Doam-myeon, Pyeongchang-gun, in 2006. Based on the sequence analysis of PCR-amplified 16S ribosomal DNA and 16S-23S spacer region DNA products using universal phytoplasma primers, the phytoplasma associated with leafy lespedeza witches' broom (LLWB) disease was identified as a member of Candidatus Pytoplasma trifolii. It was most closely related to alsike clover proliferation phytoplasma (99.8% similarity, accession no. AY390261), Candidatus Pytoplasma trifolii strain. RFLP patterns generated with AluI, HpaII clearly differentiated LLWB phytoplasma from the referenced phytoplasma strains, water dropwort witches' broom, mulberry dwarf, glehni aster yellow dwarf and jujube witches' broom. This paper is the first report on Candidatus Phytoplasma trifolii in leafy lespedeza identified at a molecular level.

Positive Association Between miR-499A>G and Hepatocellular Carcinoma Risk in a Chinese Population

  • Zou, Hong-Zhi;Zhao, Yan-Qiu
    • Asian Pacific Journal of Cancer Prevention
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    • v.14 no.3
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    • pp.1769-1772
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    • 2013
  • A case-control study of the association of miR-499A>G rs3746444 with risk of hepatocellular carcinoma (HCC)was conducted. Patients with HCC and healthy control subjects were recruited for genotyping of miR-499A>G using duplex polymerase-chain-reaction with confronting-two-pair primer(PCR-RFLP) analysis. The MiR-499 GG genotype was associated with a decreased risk of HCC as compared with the miR-499 AA genotype (adjusted OR=0.74, 95%CI=0.24-0.96). Similarly, the GG genotype showed a 0.45-fold decreased HCC risk in a recessive model. The MiR-499 G allele was significantly associated with decreased risk of HCC among patients infected with HBV in a dominant model (OR=0.09, 95%CI= 0.02-0.29). In conclusion, the MiR-499A>G rs3746444 polymorphism is associated with HCC risk in the Chinese population, and may be useful predictive marker for CAD susceptibility.

Investigating the Frequency of the ERCC1 Gene C8092A Polymorphism in Iranian Patients with Advanced Gastric Cancer

  • Mokmeli, Sharareh;Tehrani, Golnaz Asaadi;Zamiri, Reza Eghdam;Bahrami, Tayyeb
    • Asian Pacific Journal of Cancer Prevention
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    • v.17 no.3
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    • pp.1369-1372
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    • 2016
  • Background: Platinum compounds are the main drugs for treatment of advanced gastric cancer. Previous studies have shown that clinical outcome with platinum-based compounds depends on ERCC1 polymorphisms. The aim of this study was to investigate the frequency of a common polymorphism of ERCC1 gene (C8092A) in Iranian patients with advanced gastric cancer receiving platinum chemotherapy. Materials and Methods: Genetic analysis of the ERCC1 C8092A polymorphism was performed by the PCR - RFLP method using 50 paraffin-embedded tissue specimens. Results: Of the 50 cases, 32% of individuals showed CC genotype, 24% of them had CA genotype and 44% of patients had AA genotype. Conclusions: Based on the results, using of platinum-based chemotherapy would be expected to be specifically beneficial in only 32% of patients.

Molecular Cloning And analysis of Korean Insulin Gene (한국인 인슈린 유전자의 클로닝 및 분석)

  • 김형민;한상수;고건일;손동환;전창덕;정헌택;김재백
    • YAKHAK HOEJI
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    • v.37 no.5
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    • pp.504-510
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    • 1993
  • Human insulin gene is consisted of the polymorphic region with the repeating units, the regulatory sequence, the structural gene including the intervening sequence, and 3'-flanking region. The polymerase chain reaction, which amplifies the target DNA between two specific primers, has been performed for the amplification of human insulin gene and simple one-step cloning of it into Escherichia coli. Out of 1727 nuceotides compared, only 4 sites were variable: 5'-regulatory region(G2101$\rightarrow$AGG); IVS I(T2401$\rightarrow$A); Exon II(C2411 deletion); IVS II(A2740 dejection). The variations at the G2101 and T2401 were the same as those found in one American allele. The other two variations were observed only in the specific Korean allele. And, the enzyme digestion patterns among normal, insulin dependent diabetes mellitus, and non-insulin dependent diabetes mellitus were the same. On the other hand, PCR method showed the possibility of the quickaccess for the polymorphic region in terms of the restriction fragment length of polymorphism.

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THE MICROSOMAL EPOXIDE HYDROLASE(MEPHXS) POLYMORPHISMS AND THE ASSOCIATION OF MICRONUCLEI(MN) IN THE BENZENE-EXPOSED WORKERS

  • Kim, Tae-Yon;Kwon, Seul-A;Kim, Jin-Sik;Kim, Su-Young;Lee, Young-Jun;Chung, Eun-Jung;Paek, Do-Myung;Park, Jung-Keun;Chung, Hai-Won
    • Proceedings of the Korean Society of Toxicology Conference
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    • 2002.05a
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    • pp.134-134
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    • 2002
  • In order to investigate whether genetic polymorphisms in the mEH exon3 (Tyr139His), exon4 (His113Arg) interact with the formation of MN in benzene exposed workers, the PCR-RFLP based genotyping of 76 exposed and 115 controls were performed and cytokinesis-block micronucleus (CBMN) analysis of 56 exposed and 53 controls was employed.(omitted)

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Molecular Characterization of Novel Insecticidal Cryl-Type Genes from Bacillus Thuringiensis K1

  • Li, Ming-Shun;Park, Jae-Young;Roh, Jong-Yul;Shim, Hee-Jin;Boo, Kyung-Saeng;Je, Yeon-Ho
    • Proceedings of the Korean Society of Sericultural Science Conference
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    • 2003.04a
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    • pp.72-72
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    • 2003
  • A new Bacillus thuringiensis strain (Kl), having high toxicities to Plutella xylostella and Spodoptera exigua was isolated from Korean soil sample. It was determined to belong to subsp. kurstaki (H3a3b3c) and produced bipyramidal inclusion. PCR-RFLP analysis showed that this isolate contains three novel cryl-type crystal protein genes in addition to crylAa and crylE genes. (omitted)

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The Analysis of Methylenetetrahydrofolate Reductase Mutation in Recurrent Spontaneous Abortion Associated with Hyperhomocysteinemia (Homocysteine 과다증과 관련된 반복 자연유산에서 Methylenetetrahydrofolate Reductase돌연변이에 대한 분석)

  • Nam, Yoon-Sung;Choi, Jong-Soon;Ha, Kwon-Soo;Lee, Zee-Won;Oh, Do-Yeon
    • Clinical and Experimental Reproductive Medicine
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    • v.26 no.3
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    • pp.441-445
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    • 1999
  • Objective: To analyze the methylenetetrahydrofolate reductase (MTHFR) mutation in recurrent spontaneous abortion associated with hyperhomocysteinemia. Material and Method: The blood sample of habitual aborter with high fasting homocysteine level was tested by PCR - RFLP method. Results: The patient was found to be a homozygosity for MTHFR gene mutation that was confirmed by the finding which is consistent with the mutation at the nucleotide 677 C to T, corresponding to Ala to Val. Conclusions: Hyperhomocysteinemia due to MTHFR mutation is a cause of recurrent spontaneous abortion. Therefore, the MTHFR mutation should be examined in the workup of recurrent spontaneous abortion showing hyperhomocysteinemia.

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Phytoplasma Infection could Affect Chemical Composition of Artemisia sieberi

  • Hemmati, Chamran;Nikooei, Mehrnoosh
    • The Plant Pathology Journal
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    • v.35 no.3
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    • pp.274-279
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    • 2019
  • Artemisia sieberi showing symptoms resembling those caused by phytoplasma were observed in Geno Mountain, Hormozgan Province, Iran, and were examined for phytoplasma presence by PCR assays. In addition, the essential oils hydrodistilled from the aerial parts of phytoplasma-infected and healthy plants have been analyzed and compared by GC and GC/MS. Phylogenetic and virtual RFLP analysis of the 16S rRNA gene sequences revealed that the phytoplasma associated with A. sieberi witches' broom (AsWB) was a strain of 'Candidatus Phytoplasma aurantifolia'. The presence of the disease, however, induced a further enrichment (from 4.9 to 45.2%, a relative increase of 90%) of the entire monoterpene class as compared to the abundance in healthy samples. Conversely, a matching decrease in monoterpenoid (from 48.7 to 2%, a relative decrease of 90.2%) was observed in the infected plants. Besides the first report of phytoplasma infection of A. sieberi, the changes of its essential oils are reported.