• Title/Summary/Keyword: G-S PCR

Search Result 719, Processing Time 0.032 seconds

Association between p53 Gene Variants and Oral Cancer Susceptibility in Population from Gujarat, West India

  • Patel, Kinjal R.;Vajaria, Bhairavi N.;Begum, Rasheedunnisa;Shah, Franky D.;Patel, Jayendra B.;Shukla, Shilin N.;Patel, Prabhudas S.
    • Asian Pacific Journal of Cancer Prevention
    • /
    • v.14 no.2
    • /
    • pp.1093-1100
    • /
    • 2013
  • Background: p53 gene variants i.e. 16 bp duplication in intron 3, Arg72Pro in exon 4 and G>A in intron 6 have been reported to modulate susceptibility to various malignancies. Therefore, the present study evaluated the role of these p53 polymorphisms in oral cancer susceptibility in a population from Gujarat, West India. Method: Genotype frequencies at the three p53 loci in 110 controls and 79 oral cancer cases were determined by the PCR-RFLP method. Results: Heterozygous individuals at exon 4 showed protection from developing oral cancer. Homozygous wild and heterozygous individuals at intron 3 and those heterozygous at exon 4 in combination appeared to be at lowered risk. Furthermore, carriers of the 16 bp duplication allele at intron 3, proline allele at exon 4 and G allele at intron 6 were protected from oral cancer development. Conclusion: p53 polymorphisms, especially Arg72Pro in exon 4 could significantly modify the risk of oral cancer development in Gujarat, West Indian population.

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
    • /
    • v.15 no.1
    • /
    • pp.61-67
    • /
    • 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.

The Inhibitiory Effects of the Scirpi rhizoma on Melanogenesis (삼릉추출물의 멜라닌 생성 저해 효과)

  • Lee, Bum-Chun;Kim, Jin-Hwa;Sim, Gwan-Sub;Zhang, Yong-He;Pyo, Hyeong-Bae
    • Journal of the Society of Cosmetic Scientists of Korea
    • /
    • v.31 no.4 s.54
    • /
    • pp.305-310
    • /
    • 2005
  • To obtain effective and safe depigmenting agents, we investigated the effects of Scirpi rhizoma, a medicine among Chinese herbs, on melanogenesis. Dried S. rhizoma was refluxed with 70% aqueous ethanol and the extract was evaporated to dryness. To determine the effects as a whitening agent, various in vitro tests were performed such as free radical scavenging activity, melanin formation assay, tyrosinase activity and expression of tyrosinase, TRP-1 and TRP-2(western blot and RT-PCR) in B16 melanoma cells. S. rhizoma showed scavenging activities of free radicals and reactive oxygen species (ROS) with the $IC_{50}\;of\;638{\mu}g/mL$ against 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical and $21.7{\mu}g/mL$ against superoxide radicals in the xanthine/xanthine oxidase system, respectively. S. rhizoma significantly inhibited melanin production in B16 melanoma cells. S. rhizoma treatment(48 h) suppressed the biosynthesis of melanin up to 27% at 100{\mu}g/mL$ and reduced tyrosinase activity up to 31% at $100{\mu}g/mL$ in B16 melanoma cells. S. rhizoma was also able to significantly inhibit tyrosinase and TRP-1 expression in protein and mRNA level. These results suggest that S. rhizoma inhibited melanin biosynthesis by regulating tyrosinase activity and expression in B16 melanoma cells. Therefore, S. rhizoma may be useful as a new antioxidant and whitening agent to inhibit melanogenesis.

Analysis of the GCK and HNF-1α Gene Polymorphism in Korean Type 2 Diabetic Patients by PCR-DHPLC (PCR-DHPLC를 이용한 한국인 제2형 당뇨환자의 GCK와 HNF-1α의 유전자다형성 분석)

  • Nam, Youn-Hyoung;Park, Dae-Yong;Park, Sang-Bum;An, Young-Chang;Lee, Sang-Hyun;Cho, Min-Ho;Park, Su-Min;Jang, Won-Cheoul
    • Journal of the Korean Chemical Society
    • /
    • v.51 no.6
    • /
    • pp.543-548
    • /
    • 2007
  • Maturity-onset diabetes of the young (MODY) is a genetically heterogeneous subtype of Type 2 diabetes characterized (non-insulin-dependent) by early onset, usually before 25 years of age, autosomal dominant inheritance and a primary defect in insulin secretion. Mutations in the glucokinase (GCK) and hepatocyte nuclear factor (HNF)-1α genes are the major causes of monogenic forms of Type 2 diabetes mellitus. Therefore it is need to study relation with these polymorphisms by diverse analysis methods. The promotor and coding regions inclusive intron exon boundaries of the GCK, HNF-1α genes were examined by PCR-DHPLC (Polymerase Chain Reaction - Denaturing High Performance Liquid Chromatography) and direct sequencing. We extracted DNA from 11 patients and 20 normals. Then we confirmed a single-nucleotide polymorphism using PCR-DHPLC. As results, we identified one mutation (R135G) in GCK gene and two mutations (I27L, S487N) in HNF-1a and at the same time detected mutation in intron 8.

Detection of Toxoplasma gondii in experimentally infected porcine blood and tissues by polymerase chain reaction (Polymerase chain reaction을 이용한 실험적 감염 돼지의 혈액과 조직으로부터 Toxoplasma gondii 검출)

  • Suh, Myung-deuk;Shin, Gee-wook
    • Korean Journal of Veterinary Research
    • /
    • v.41 no.1
    • /
    • pp.89-98
    • /
    • 2001
  • This study was conducted to detect the toxoplasma specific-DNA in circulating blood and organs collected from slaughtered pigs at slaughtering house and experimentally infected pigs with Toxoplasma gondii tachyzoites by polymerase chain reaction(PCR), and also PCR was applied to diagnose for acute phase of swine toxoplasmosis as a newly developed diagnostic test. The sensitivity of oligonucleotide primer, T-1 & T-2, designed from toxoplasma B1 gene amplification method was compared with Tp parasite detection by mouse inoculation(MI). On the other hand, latex agglutination test(LAT) was conducted to detect the serum antibodies comparing with the detection of toxoplasma by PCR and MI. The results obtained were summarized as follows. PCR was able to determine at the lowest level of $10^0/ml$ T. gondii in blood samples which were blended with a serial diluted T gondii in vitro. On the other hand, $10^2/5g$ of T gondii could detect from a variety of tissues including lung, diaphragm, liver, heart, spleen and brain in vitro. The primer was proved to specifically determine T gondii in blood and tissues in vitro but it did not detect Neospora caninum used as a negative control. DNA of T. gondii was effectively extracted by freezing, thawing and grinding twice both tissues mixed with T gondii in vitro and in experimentally infected pig's tissues. PCR detected specific DNA in the blood of experimentally infected pigs at 108 hrs and 120 hrs post-infection, it was the same time that the pigs showed fever and parasitaemia. In case of tissue, specific DNA was, however, detected only lung from experimentally infected pigs. Even though the duration of acute phase was from 3 to 7 days post-infection, but the latex agglutination test (LAT) results appeared from 8 days post-infection. A comparison of sensitivity in determining T gondii in blood samples between PCR and MI, PCR positive rate ranged from 25 to 33.3%, but that of MI covered from 75 to 100%.

  • PDF

Pathogenic and Molecular Characteristics of Agrobacterium vitis strains isolated from Grapevine in Korea

  • Kim, J.G.;Kim, S.H.;Choi, J.E.;Lee, Y.K.;Kang, H.W.
    • Proceedings of the Korean Society of Plant Pathology Conference
    • /
    • 2003.10a
    • /
    • pp.120.2-120
    • /
    • 2003
  • Agrobacterium vitis is a causal agent of crown-gall disease on grapevine. In Korea, grapevine variety (GeoBong) have severely been infected by the bacteria since stems of the variety were buried in soil for overwintering. Infection ratio over 70-80% was observed on 7 years old GeoBong grapevine in Ansung and Cheonan. PCR specific primers for A. vitis strains were designed using nucleotide sequences of vir A gene in Ti-Plasmid, pheA gene in chromosomal DNA and a URP-PCR polymorphic band. Three hundred bacterial strains were isolated from the different 80 galls formed on GeoBong grapevine in Cheonan and Ansung of Korea and were screened to identify A. vitis using the three specific PCR primers for Agrobacterium vitis. Twenty-four bacterial strains that are detected by the primers were further confirmed by pathogenicity and biochemical methods. To investigate the genomic diversity of the bacterial strains, twenty primers of 20 mer referred to universal rice primers (URP) were applied for PCR fingerprinting, Of them, URP2R and URP2F primers could effectively be used to detect polymorphism within the bacterial strains.

  • PDF

A RAPD-PCR Method for the Rapid Detection of Bacillus cereus

  • Lee, Ji-Yeon;Kwon, Gun-Hee;Park, Jae-Yong;Park, Cheon-Seok;Kwon, Dae-Young;Lim, Jin-Kyu;Kim, Jong-Sang;Kim, Jeong-Hwan
    • Journal of Microbiology and Biotechnology
    • /
    • v.21 no.3
    • /
    • pp.274-276
    • /
    • 2011
  • Distinction of Bacillus cereus from other closely related bacilli is challenging and new efficient methods are continually demanded. From our previous work on RAPD profiles of bacilli, we found a possibility that B. cereus strains could be distinguished from other bacilli. In this work, RAPD-PCR profiles of B. cereus strains were obtained using a 10-mer (S30) as a primer, and a B. cereus specific 0.91-kb band was produced from all tested strains. The RAPD-PCR procedure also successfully detected B. cereus from spiked cheonggukjang when B. cereus cells were present at more than $10^2$/g sample.

Effects on Hot Water Extract of Schizandra chinensis on Colon Cancer (오미자 열수추출물의 대장암세포 증식억제 효과)

  • Ryu, Min-Ju;Chung, Ha-Sook
    • Food Engineering Progress
    • /
    • v.15 no.1
    • /
    • pp.64-69
    • /
    • 2011
  • The anticancer activity of Schizandra chinensis Baillon was investigated for the development of functional food resources. The antiproliferative activity of hot water extracts of Schizandra chinensis Baillon in human colon cancer cell line (HT-29) were identified using cell viability, morphology study, cell cycle and RT-PCR analyses. HT-29 cells were cultured in several concentrations (0, 1.0, 2.0, 4.0 mg/mL) of water extracts of Schizandra chinensis Baillon. In our study, colon cancer cell growth could be inhibited by hot water extracts of Schizandra chinensis Baillon in a dose-dependent manners. It was associated with morphological changes and apoptotic cell death with cell shrinking, chromatin condensation, apoptotic bodies and cell cycle analysis. These results suggest that Schizandra chinensis Baillon may inhibit the growth of human colon cancer cells by various apoptosis-aiding activities as well as apoptosis itself.

Analysis of Foodborne Pathogens in Brassica campestris var. narinosa microgreen from Harvesting and Processing Steps (어린잎채소의 생산 및 가공 공정 중 식중독 미생물 분석)

  • Oh, Tae Young;Baek, Seung-Youb;Choi, Jeong Hee;Jeong, Moon Cheol;Koo, Ok Kyung;Kim, Seung Min;Kim, Hyun Jung
    • Journal of Applied Biological Chemistry
    • /
    • v.59 no.1
    • /
    • pp.63-68
    • /
    • 2016
  • This study was performed to assess the microbiological quality of Brassica campestris var. narinosa microgreen from harvesting and processing steps. The samples were analyzed for total viable cell counts (TVC), coliforms, Enterobacteriaceae, Escherichia coli, Salmonella spp., Listeria monocytogenes, Vibrio parahaemolyticus, Bacillus cereus, and Staphylococcus aureus. The total viable counts of microgreen (whole leaves) and environment samples from harvesting steps were higher than 6.8 log CFU/g and the contamination level of coliforms in the samples were 3.2 log CFU/g and 3.5 log CFU/g of microgreen and soil, respectively. In case of microgreen samples collected from processing steps, the contamination level of TVC and coliforms were higher in raw materials than samples obtained from later stages of processing, i.e. washing, drain, and final products. The contamination levels of B. cereus in raw materials and environments decreased approximately 1.4 log CFU/g in final products. S. aureus was detected in soil samples but Salmonella spp., Listeria monocytogenes, Vibrio parahaemolyticus and pathogenic E. coli was not detected. In order to identify the sources of contamination for microgreen, the genetic similarity of B. cereus isolates obtained from harvesting and processing steps were compared using the repetitive-sequence-based polymerase chain reaction method. B. cereus isolates obtained from harvesting environments and microgreen were clustered with a similarity greater than 95%. In case of B. cereus isolates obtained from microgreen and environmental samples at processing steps showed low genetic similarity.

Phylogenetic Analysis by RFLP and Sequencing of Mitochondrial DNA in a Korean Population

  • Lee, Jin-Young;Kim, Heui-Soo;Ha, Bae-Jin;Park, Yeong-Hong
    • Archives of Pharmacal Research
    • /
    • v.29 no.1
    • /
    • pp.88-95
    • /
    • 2006
  • Analysis of molecular nature of mitochondrial DNA (mtDNA) could be powerful marker for anthropological studies of modern populations. While population genetic studies on mtDNA have been reported for several ethnic groups, no such study has been documented for the Korean population. We surveyed mtDNA polymorphisms in the HVS I of noncoding D-loop region and its upstream region from 430 unrelated healthy Korean population by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and direct sequencing analysis. PCR product with 2,790 bp spanning the specific mtDNA region (mt13715-16504) was subjected to RFLP analysis using 6 restriction enzyme (Hinf I, Hae III, Alu I, Dde I, Mbo I, Rsa I). On the PAUP analysis of PCR-RFLP results, 38 mtDNA haplotypes (Hap 1-38) were detected in the Korean populations, which were classified into 11 haplogroups (Grp 1-11) of related haplotypes encompassing all 38 haplotypes. In comparison of sequencing data with Anderson's reference sequence, the transition type was more prevalent than the transversion type. Insertions or deletions were not found. In addition, three of the polymorphic sites (A16240C, A16351G, G16384A) in HVS-I region are determined newly. The polymorphic sites were distributed randomly in the region, though the frequency at each site was variable. Thus, this research might be required for the genealogical study of Orientals.