• Title/Summary/Keyword: Point mutation

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Cross Resistance and Point Mutation of the Mitochondrial Cytochrome b of Bifenazate Resistant Two-spotted Spider Mite, Tetranychus urticae (Bifenazate 저항성 점박이응애에 대한 교차저항성과 미토콘드리아 cytochrome b의 점돌연변이)

  • Lee, Kyo-Ri;Koo, Hyun-Na;Yoon, Chang-Mann;Kim, Gil-Hah
    • The Korean Journal of Pesticide Science
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    • v.14 no.3
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    • pp.247-254
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    • 2010
  • Two-spotted spider mite, Tetranychus urticae was collected from the rose greenhouse in Chilgok, Gyeongbuk Province in December 2000. This population has been selected for ten years with bifenazate (over 450 times), and increased 855.9 fold in resistance as compared with susceptible strain (S). Cross resistance of bifenazate resistant (BR) strain to eight miticides was investigated. The BR strain exhibited high and low cross resistance to acequinocyl (614.0 fold) and to chlorfenapyr (9.1 fold), respectively. Against fenazaquin (0.3 fold) and fenpyroximate (0.1 fold), however, showed the strain negatively correlated cross resistance. Each strain collected in Choeng-ju (CJ), Kang-jin (KJ), and Chung-ju (CUJ) showed 5.5-, 964.5-, and 21.8-fold resistance to bifenazate, respectively. The detoxifying enzymes of the BR strain showed 1.6-fold activity in cytochrome $P_{450}$-dependent monooxygenase ($P_{450}$) as compared with susceptible one. By comparing the mitochondrial cytochrome b (cytb) sequence, G126S point mutation was detected in the BR and KJ strains.

ACTIVATION OF H-RAS ONCOGENE IN RAT SALIVARY GLAND TUMORS INDUCED BY DMBA AND IRRADIATION (DMBA 매식과 방사선 조사로 유도된 백서 타액선 종양에서 H-ras 암유전자의 활성화)

  • Hu Key-Soon;Choi Jong-Whan;Choi Soon-Chul;Park Tae-Won;You Dong-Soo
    • Journal of Korean Academy of Oral and Maxillofacial Radiology
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    • v.28 no.1
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    • pp.245-259
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    • 1998
  • Cellular transforming genes have been identified in a number of different tumor cell lines and tumor types. A significant number of these oncogenes belong to the ras gene family. The ras gene family consists of three closely related genes:H-ras, K-ras and N-ras which code for a related 21 kDa protein. Mutations in codon 12, 13 and 61 of one of the three ras genes convert these genes into acute oncogenes. The presence of H-ras gene mutations has important prognostic implications in various tumors. Each genomic DNA was isolated from tumors induced by implantation with DMBA, or by treatment with DMBA -implantation/irradiation. When genome DNA was transfected into NIH 3T3 cells and investigated by two-step PCR-RFLP, the fOllowing results were concluded: 1. Transformation foci developed in two groups when the genome DNA of two experimental groups were transfected into NIH 3T3 cells. 2. Transformation efficiency was 0.01-0.02 foci/㎍DNA in the experimental group with the DMBA-implantation, 0.01-0.03 foci/㎍lgDNA in the experimental group with the DMBA-implantation/irradiation according to results of transfection assay. 3. When the point mutation of H-ras gene was investigated by a two-step PCR-RFLP, there was 13.9% (5/36) in the experimental group with the DMBA implantation, 15.4 % (6/39) in the experimental group with the DMBA -implantation/irradiation. 4. The point mutation in codon 12 and 61 of H-ras was 5.6%(2/36) and 8.3%(3/36) in the experimental group with the DMBA implantation. 5. The point mutation in codon 12 and 61 of H-ras gene was 7.7%(3/39) in the experimental group with the DMBA -implantation/irradiation.

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Monitoring of Bifenazate Resistant Two-spotted Spider Mite, Tetranychus urticae Using Molecular Detection Method (분자학적 진단방법을 이용한 bifenazate 저항성 점박이응애 모니터링)

  • Lee, Kyu-Ri;Shin, Yun-Ho;Cho, Sun-Ran;Koo, Hyun-Na;Choi, Jang-Jeon;Ahn, Ki-Su;Kim, Gil-Hah
    • The Korean Journal of Pesticide Science
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    • v.15 no.1
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    • pp.61-67
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    • 2011
  • In 2010, two-spotted spider mite, Tetranychus urticae was collected from the rose greenhouse and apple orchards in Cheongju (CJ), Chungju (CUJ)-1, CUJ-2, Kangjin (KJ), Yesan (YS), and Yeongju (YJ). Among them, KJ and YS strain showed high resistance to bifenazate of 964.5- and 1l30-fold, respectively. The other strains showed low resistance to bifenazate. By analyzing the mitochondrial cytochrome b (cytb) sequence, G126S point mutation was detected in KJ and YS strain. Thus, G126S point mutation in the mitochondrial cytb was available molecular detection marker for selection of bifenazate resistant T. urticae. Two molecular detection methods, quantitative sequencing (QS) and PCR amplification of specific alleles (PASA) were well detected specific G126S point mutation. Therefore, these methods can be used to monitor the resistance allele in field population of T. urticae and bifenazate resistance management strategy.

Study on the expression and detection of the p53 mutation in Korean colon cancer cell lines (한국인의 대장암 세포주에서 p53 돌연변이의 발견과 발현에 관한 연구)

  • Jung, Ji-Yeon;Oh, Sang-Jin
    • IMMUNE NETWORK
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    • v.1 no.2
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    • pp.151-161
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    • 2001
  • Background: Inactivation in p53 tumor suppressor gene through a point mutation and deletion is one of the most frequent genetic changes found in human cancer, with 50% of an incidence. This high rate of mutation mostly suggests that the gene plays a central role in the development of cancer and the mutations detected so far were found in exons 5 to 8. Mutation of p53 locus produced accumulation of abnormal p53 protein, and negative regulation of cell proliferation and transcriptional activation as a suppressor of transformation were lost. In addition, inhibition of its normal cellular function of wild-type by mutant is an important step in tumorigenesis. Method: 4 colon cancer cell lines (SNU C1, C2A, C4, C5) were examined for mutation in exons 5 to 8 of the p53 tumor suppressor gene by PCR-SSCP analysis and expression pattern by western blotting and immunoprecipitation. p53-mediated transactivation ability were examined by CAT assay and base substitution of p53 in SNU C2A cell were detected by DNA sequencing. Results: 1) SNU C2A cell and SNU C5 cell were detected mobility shifts each in exon 5 and exon 7 of p53 gene by the PCR-SSCP method, implicating being of p53 mutation. 2) 3 colon cancer cell lines (SNU C1, SNU C2A, SNU C5) expressed wild type and mutant type p53 protein. 3) In northern blot experiment, SNU C2A and SNU C5 cell expressed high level of p53 mRNA. 4) Results of p53-mediated transactivation in colon cancer cell lines by CAT assay represented only SNU C2A cell has transcriptional activity. 5) DNA sequencing in SNU C2A cell showed missense mutation in codon 179 of one allele, histidine to arginine and wild type p53 in the other allele. Conclusion: Colon cancer cell lines showed correlation with mutation in p53 gene and accumulation of abnormal p53 protein. Colon cancer cell SNU C2A retained p53-mediated transactivation as heterozygous p53 with one mutant allele in 179 codon and the other wild-type allele.

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Hyperthyroidism Caused by a Mutation in the Thyrotropin Receptor Gene in Two Brothers (형제에서 발생한 갑상선 자극호르몬 수용체 유전자 돌연변이에 의한 갑상선 항진증 2례)

  • Kim, Jae Hyun;Lee, Sung Soo;Lim, Jung Sup;Shin, Choong Ho;Yang, Sei Won
    • Clinical and Experimental Pediatrics
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    • v.48 no.3
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    • pp.337-341
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    • 2005
  • Thyrotropin receptor(TSHR) mutations must be considered when congenital hyperthyroidism has persisted, but there has been no evidence for autoimmunity. TSHR mutations leading to constitutive activation of the thyroid gland were identified as the molecular cause of autosomal dominant nonautoimmune hyperthyroidism and sporadic congenital hyperthyroidism. We report two cases of hyperthyroidism caused by germline TSHR mutation who presented with exessive sweating and no evidence of autoimmune thyroid disease. They were brothers and their mother had undergone thyroidectomy because of hyperthyroidism. Direct sequencing of the polymerase chain reaction-amplified exon 10 of the TSHR genomic DNA revealed a transition of GCT to GTT, resulting in an exchange of alanine 627 to valine in the patients and their mother. This might be a novel mutation or polymorphism, but we did not perform any functional gene study. But considering the clinical profiles, we can conclude that hyperthyroidism of these two brothers might come from the point mutation described above.

The influence of p53 mutation status on the anti-cancer effect of cisplatin in oral squamous cell carcinoma cell lines

  • Jo, Deuk-Won;Kim, Young-Kyun;Yun, Pil-Young
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • v.42 no.6
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    • pp.337-344
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    • 2016
  • Objectives: The purpose of this study was to evaluate the anti-cancer activity of cisplatin by studying its effects on cell viability and identifying the mechanisms underlying the induction of cell cycle arrest and apoptosis on oral squamous cell carcinoma (OSCC) cell lines with varying p53 mutation status. Materials and Methods: Three OSCC cell lines, YD-8 (p53 point mutation), YD-9 (p53 wild type), and YD-38 (p53 deletion) were used. To determine the cytotoxic effect of cisplatin, MTS assay was performed. The cell cycle alteration and apoptosis were analyzed using flow cytometry. Western blot analysis was used to detect the expression of cell cycle alteration- or apoptosis-related proteins as well as p53. Results: Cisplatin showed a time- and dose-dependent anti-proliferative effect in all cell lines. Cisplatin induced G2/M cell accumulation in the three cell lines after treatment with 0.5 and $1.0{\mu}g/mL$ of cisplatin for 48 hours. The proportion of annexin V-FITC-stained cells increased following treatment with cisplatin. The apoptotic proportion was lower in the YD-38 cell line than in the YD-9 or YD-8 cell lines. Also, immunoblotting analysis indicated that p53 and p21 were detected only in YD-8 and YD-9 cell lines after cisplatin treatment. Conclusion: In this study, cisplatin showed anti-cancer effects via G2/M phase arrest and apoptosis, with some difference among OSCC cell lines. The mutation status of p53 might have influenced the difference observed among cell lines. Further studies on p53 mutation status are needed to understand the biological behavior and characteristics of OSCCs and to establish appropriate treatment.

JAK-2 V617F Mutational Analysis in Primary Idiopathic Myelofibrosis: Experience from Southern Pakistan

  • Sultan, Sadia;Irfan, Syed Mohammed
    • Asian Pacific Journal of Cancer Prevention
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    • v.16 no.17
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    • pp.7889-7892
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    • 2015
  • Background: Primary idiopathic myelofibrosis (PMF) is a clonal Ph-chromosome negative myeloproliferative neoplasm characterized by dysregulated kinase signaling and release of abnormal cytokines. In the recent past, following JAK2 V617F mutation invention, important revolution has been made in the molecular diagnostic biology of this disease. The rational of this study was to determine the mutational status of JAK2 V617F in Pakistan patients with PMF. Materials and Methods: In this cross sectional study, 20 patients with PMF were enrolled from January 2011 to December 2014. Diagnosis was based on WHO criteria for PMF. All patients were screened for G-T point mutation (V617F) in the JAK2 gene on chromosome 9 by allele specific PCR. Results: The mean age was $57.9{\pm}16.5years$. The male to female ratio was 3:1. The frequency of JAK2 V617F positivity in our PMF patients was found to be 55%. Positive correlations of JAK2 V617F mutation were established with high TLC count, raised LDH and marked splenomegaly (P<0.05). No correlation of JAK2 V617F could be established with age and gender (P>0.05). Conclusions: The JAK2 V617F mutation frequency in our PMF patients was similar to those reported previously. In our hands JAK2 V617F mutated patients expressed an aggressive disease phenotype. Screening for the mutation in all suspected PMF cases could be beneficial in differentiating patients with reactive and clonal marrow fibrosis.

Resistance Monitoring and Analysis of Point Mutations to λ-cyhalothrin, Imidacloprid, and Flupyradifurone in Field-collected Populations of Myzus persicae (Hemiptera: Aphididae) (복숭아혹진딧물 야외개체군의 λ-cyhalothrin, imidacloprid, 그리고 flupyradifurone에 대한 저항성 모니터링과 점 돌연변이 분석)

  • Ha Hyeon Moon;Yuno Lee;Dong-Hyun Kang;Se Eun Kim;Hyun Kyung Kim;Hyun-Na Koo;Gil-Hah Kim
    • Korean journal of applied entomology
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    • v.63 no.1
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    • pp.33-42
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    • 2024
  • The green peach aphid, Myzus persicae is a representative agricultural insect pest that is polyphagous and causes serious damage to tobacco, potatoes, peppers, cabbage, and peaches. In this study, we analyzed the level of development of insecticide resistance to λ-cyhalothrin, imidacloprid, and flupyradifurone and the point mutations (R81T, L1014F, M918L) in 12 field populations of M. persicae. In addition, the expression level of CYP6CY3, a cytochrome P450 gene, was analyzed through qRT-PCR. As a result, λ-cyhalothrin showed high resistance ratio (RR) of > 200 in all 12 populations. Imidacloprid and flupyradifurone showed high RR of >200 in YS, UR, HY, and WJ populations. The R81T was detected in approximately 50%, L1014F in approximately 33.3%, and M918L in 100% of the 12 populations. Additionally, the expression level of subunit CYP6CY3 was highest in imidacloprid-resistant population (YS). These results suggest that M918L point mutation can be used as λ-cyhalothrin-resistance molecular diagnostic and R81T point mutation and the high expression of CYP6CY3 can be used as imidacloprid-resistance molecular diagnostic markers.

Chimeric Protein of CD8a Extracellular Domain and CD4 Transmembrane and Cytoplasmic Domain Binds More Efficiently to p561ck than CD8a

  • Young Il Choi;Sang Dai Park;Rho Hyun Seong
    • Animal cells and systems
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    • v.3 no.3
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    • pp.331-336
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    • 1999
  • p56$^{Ick}$, a cytoplasmic protein tyrosine kinase of the src family, is non-covalently associated with the cell surface coreceptors CD4 and CD8, which are expressed on thymocytes and mature T cells. The coreceptor protein plays an important role during the differentiation of thymocytes and the activation of T cells. DNA constructs were designed to study the roles of CD4 and CD8 during the differentiation of thymocytes. One is a chimeric cDNA which consists of coding regions for the extracellular domain of CD8a and the transmembrane and cytoplasmic domain of CD4. The other is the same chimeric cDNA but with a point mutation converting Cys to Ala in the Ick-binding site to disrupt the association. We confirmed that the CD8a/CD4 chimeric molecule bound to Ick more efficiently than the wild type CD8a protein. However, the chimeric protein with the Cys$leftrightarro$Ala mutation did not associate with Ick. The results suggest a possibility that the CD8a/CD4 chimeric protein may behave like a CD4 protein in associating with Ick and that it may deliver a signal inside the cell in a similar manner, Analysing effects of the mutant CD8a/CD4 chimeric protein expression in developing thymocytes will elucidate the role of Ick during the determination of CD4/CD8 cell lineages.

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Machine Learning Perspective Gene Optimization for Efficient Induction Machine Design

  • Selvam, Ponmurugan Panneer;Narayanan, Rengarajan
    • Journal of Electrical Engineering and Technology
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    • v.13 no.3
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    • pp.1202-1211
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    • 2018
  • In this paper, induction machine operation efficiency and torque is improved using Machine Learning based Gene Optimization (ML-GO) Technique is introduced. Optimized Genetic Algorithm (OGA) is used to select the optimal induction machine data. In OGA, selection, crossover and mutation process is carried out to find the optimal electrical machine data for induction machine design. Initially, many number of induction machine data are given as input for OGA. Then, fitness value is calculated for all induction machine data to find whether the criterion is satisfied or not through fitness function (i.e., objective function such as starting to full load torque ratio, rotor current, power factor and maximum flux density of stator and rotor teeth). When the criterion is not satisfied, annealed selection approach in OGA is used to move the selection criteria from exploration to exploitation to attain the optimal solution (i.e., efficient machine data). After the selection process, two point crossovers is carried out to select two crossover points within a chromosomes (i.e., design variables) and then swaps two parent's chromosomes for producing two new offspring. Finally, Adaptive Levy Mutation is used in OGA to select any value in random manner and gets mutated to obtain the optimal value. This process gets iterated till finding the optimal value for induction machine design. Experimental evaluation of ML-GO technique is carried out with performance metrics such as torque, rotor current, induction machine operation efficiency and rotor power factor compared to the state-of-the-art works.