• Title/Summary/Keyword: PCR polymorphism

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Association of glycophorin A mutant frequency and urinary PAH metabolites influenced by genetic polymorphisms of GSTM1 in incineration workers (소각장 근로자에서 GSTM1의 유전자 다형성이 glycophorin A변이 발현율과 소변내 PAH 대사산물 농도와의 관계에 미치는 영향)

  • 이경호;하미나;최재욱;조수헌;박정규;황응수;강대희
    • Environmental Mutagens and Carcinogens
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    • v.21 no.2
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    • pp.149-155
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    • 2001
  • Eighty-one workers including 38 employees directly incinerating industry wastes were recruited from a company located in South Korea. To evaluate the association between urinary 1-hydroxypyrene glucuronide (1-OHPG) levels, as internal dose of polycyclic aromatic hydrocarbon (PAH) exposure, and glycophorin A (GPA) mutation frequency, as an early biologic effect indicator. Urinary 1-OHPG levels were measured by synchronous fluorescence spectroscopy after immunoaffinity purification using monoclonal antibody 8E11. Erythrocyte GPA variant frequency (NN or NO) was assessed in MN heterozygotes with a flow cytometic assay. The GSTM1 and GSTT1 genotypes were assessed by a multiplex PCR method. The GPA NN phenotype frequency was higher in occupationally exposed group (n=14, mean$\pm$S.D. 6.6$\pm$12.0 in 10/SUP 6/ erythrocyte cells) than in non-exposed group (n=22, 2.1$\pm$3.5). Similarly, the GPA(NO or NN) phenotype frequency was higher in exposed group (n=14, 9.7$\pm$17.3) than non-exposed group (n=22, 4.2$\pm$6.3). The above differences failed to reach statistical significance, but a significant increase was seen in GPA variant frequency levels with increase in urinary 1-OHPG levels (Spearman's correlation: p=0.06 (NO), p=0.07 (NO or NN)). When this association was evaluated by GSTM1 genotype status, the association between GPA mutation and urinary 1-OHPG levels was stronger in individuals with GSTM1 present genotype (Spearmans correlation; r=0.50, p=0.02). These results suggest that the association between urinary 1-OHPG and GPA mutation is be modulated by the GSTM1 genotype.

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Polymorphism of Exon 2 of BMP15 Gene and Its Relationship with Litter Size of Two Chinese Goats

  • Wang, Yuqin;Yuanxiao, Li;Nana, Zhang;Zhanbin, Wang;Junyan, Bai
    • Asian-Australasian Journal of Animal Sciences
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    • v.24 no.7
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    • pp.905-911
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    • 2011
  • Polymorphisms of BMP15 gene exon 2 and its relationship with prolificacy of goats were detected by PCR-SSCP and DNA sequencing methods in Chinese two local goat breeds. The results showed that the product amplified by the primers displayed polymorphisms. Three genotypes (AA, BB and AB) were detected in Funiu white goats, and their frequency was 0.071, 0.715, 0.214, respectively. Two genotypes (AB and BB) were detected in Taihang black goats, and their frequency was 0.342 and 0.658, respectively. Sequencing revealed that four mutations (456T${\rightarrow}$G, 466C${\rightarrow}$G, 510C${\rightarrow}$T, 511T${\rightarrow}$C) occurred in genotype BB of Funiu white goat, which resulted in amino acid substitution of V155G and S171P. No mutation was detected in Taihang black goat. The Funiu white goat with genotype BB had 0.91 or 0.82 kids, more than those with AB or AA, respectively. The difference of the least squares means for litter size between BB and AB was not significant (p>0.05) in Taihang black goat. It is concluded that the BMP15 gene may be a major gene which affects the prolificacy in Funiu white goats. This study could provide basic molecular data on the reproductive characteristics of local breeds of Henan province in China, and a scientific basis for the conservation and utilization of those two goat breeds.

The effect of CYP1A2 gene polymorphisms on Theophylline metabolism and chronic obstructive pulmonary disease in Turkish patients

  • Uslu, Ahmet;Ogus, Candan;Ozdemir, Tulay;Bilgen, Turker;Tosun, Ozgur;Keser, Ibrahim
    • BMB Reports
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    • v.43 no.8
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    • pp.530-534
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    • 2010
  • Cytochrome P450 (CYP) 1A2 gene polymorphisms are thought to be involved in the metabolism of theophylline (TP). We aimed to investigate the effect of CYP1A2*1C, CYP1A2*1D, CYP1A2*1E, and CYP1A2*1F polymorphisms of the CYP1A2 on TP metabolism by PCR-RFLP in 100 Turkish patients with chronic obstructive pulmonary disease (COPD) receiving TP. One hundred and one healthy volunteers were included as control group. The genotype frequencies of the CYP1A2*1D and CYP1A2*1F were found to be significantly different in the patients compared to the controls. The "T" allele at -2467 delT and the "C" allele at -163 C > A in the CYP1A2 displayed association with a significantly increased risk for COPD. "T" allele at -2467 delT was also associated with a high risk of disease severity in COPD. In conclusion, our data suggest that genetic alterations in CYP1A2 may play a role both in the pharmacogenetics of TP and in the development of COPD.

The PGC-II Polymorphism of the Peroxisome Proliferator-activated Receptor ${\gamma}$ Coactivator $1{\alpha}$ (PGC-$1{\alpha}$) Gene in Korean Subjects with the Metabolic Syndrome

  • Im, Sun-Ok;Kim, In-Sik;Kang, Sang-Sun;Hyun, Sung-Hee
    • Biomedical Science Letters
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    • v.18 no.2
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    • pp.139-145
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    • 2012
  • RFLP of PGC-$1{\alpha}$ gene of 285 Korean women was analyzed by PCR and HpaII restriction. We evaluated the correlation between PGC 1 genotypes and biochemical results, using the results of RFLP. Study subjects were divided into 3 groups: normal group (who has been average value of serum biochemical analysis), upper group (who has been higher value than average value), and low group (who have been lower value than average value). The frequencies of $H_1H_1$, $H_1H_2$, and $H_2H_2$ genotypes were 92 (32%), 85 (32%), and 108 (38%) respectively, and the ratio between $H_1$ and $H_2$ alleles was 1:1.1. There were no meaningful differences between biochemical results and PGC-$1{\alpha}$ genotypes in the normal group. But, in upper group, there was significant difference in total cholesterol (P=0.04) level. In the result of Turkey multiple comparison test, the P value of $H_1H_1$ and $H_2H_2$ was 0.059. In upper group, there were noticeable differences also in triglyceride (P=0.034) level and glucose (P=0.043) level, respectively. There were important differences between $H_1H_1$ type and $H_1H_2$ type in triglyceride (P=0.029) level and between $H_1H_2$ type and $H_2H_2$ type in glucose (P=0.040) level. This study may provide the PGC-$1{\alpha}$ genotype patterns for the amounts of lipid and glucose in the serum. $H_2$ allele (Ser482) of PGC-$1{\alpha}$ gene may be related with upper group in Korean women.

The Studies on Identification of Lophatherum gracile(淡竹葉) Using AFLP fingerprinting Methods (AFLP fingerprinting법을 이용한 담죽엽의 감별법 연구)

  • Shim, Young-Hoon;Seong, Rack-Seon;Park, Ju-Young;Kim, Ji-Yeon;Cho, Chang-Hee;Lee, Jong-Hwa;Hyeon, Seong-Ye;Kim, Sun-Ho;Kim, Dong-Sup;Jang, Seung-Yeup
    • Korean Journal of Pharmacognosy
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    • v.41 no.4
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    • pp.297-302
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    • 2010
  • Lophatheri Herba is the aerial part of Lophatherum gracile Bronghiart(淡竹葉, Gramineae). 25~75 cm in length. Stem: cylindrical with nodes, empty inside, externally pale yellowish green. Leaf: dehiscent of lanceolate lamina, shrunken and rolled, 5~20 cm long, 10~35 mm wide; surface: pale green ~ yellowish green, parallel-formed with veins of square reticulate, more distinct of appearance on the lower surface. Banbusae Caulis In Taeniam is the stringy strip derived from the stem with the peeled-off epidermis of Phllostachys nigra Munro var. henosis Stapf, and Phllostachys bambusoides Siebold et Zuccarini(竹葉, Gramineae). Irregular in size and shape, thin plane ~ strip-shaped, sometimes powdery, sometimes 1~3 mm thick. Outer surface: pale green ~ yellowish green, sometimes grayish white L. gracile and P. nigra have different origins although they show similar morphologic features. We were able to distinguish between L. gracile and P. nigra which are almost indistinguishable through this study. AFLP(Amplifide Fragment Length Polymorphism) was more suitable for identifying differences between L. gracile and P. nigra in comparison with other genetic analysis using chemical analysis. Therefore. molecular biological methods are believed to be useful for discovering origins of herbal medicines.

Potential of Cross-infection of Colletotrichum Species Causing Anthracnose in Persimmon and Pepper

  • Kim, Hye-Ryoung;Lim, Tae-Heon;Kim, Joo-Hyung;Kim, Young-Ho;Kim, Heung-Tae
    • The Plant Pathology Journal
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    • v.25 no.1
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    • pp.13-20
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    • 2009
  • Ninety isolates of Colletotrichum species from new persimmon tree twigs and 50 isolates from pepper plant fruits were isolated via single-spore isolation. Of the 140 isolates, 26 were examined for mycelial growth, carbendazim sensitivity, and ITS sequence. Four of the isolates from the persimmon trees, which were cultivated exclusively in an orchard, showed fast mycelial growth and sensitivity to carbendazim, while five of the pepper isolates showed slower mycelial growth and were resistant to the fungicide. However, 17 isolates from persimmon trees cultivated with pepper plants in the same orchard showed slow mycelial growth like the pepper isolates and they were sensitive to carbendazim like the persimmon isolates. ITS sequence analysis of these 27 isolates led to the identification of the 22 persimmon isolates as C. gloeosporioides and the five pepper isolates as C. acutatum. PCR with species-specific primers confirmed that the 90 isolates from persimmon were C. gloeosporioides whereas the 50 isolates from pepper were C. acutatum. The 90 persimmon isolates of C. gloeosporioides and 50 pepper isolates of C. acutatum were compared by a wound inoculation test to determine their capacity for host cross-infection. All of the C. acutatum isolates from pepper caused typical symptoms of anthracnose on the fruits of pepper plants and twigs of persimmon; they differed from the C. gloeosporioides isolates from persimmon, more than 90% of which were able to infect only persimmon. Amplified fragment length polymorphism analysis revealed the existence of two groups (C. gloeosporioides and C. acutatum isolates group). At 80% genetic similarity, the C. gloeosporioides group was defined within four clusters, while the C. acutatum group was within three clusters. However, these clusterings were unrelated with the virulence of Colletotrichum species against pepper fruits.

Characterization of antimicrobial resistance and application of RFLP for epidemiological monitoring of thermophilic Campylobacter spp. isolated from dogs and humans in Korea

  • Cho, Hyun-Ho;Kim, Sang-Hyun;Min, Wongi;Ku, Bok-Kyung;Kim, Jong-Hyun;Kim, Yong-Hwan
    • Korean Journal of Veterinary Research
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    • v.54 no.2
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    • pp.91-99
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    • 2014
  • An antimicrobial susceptibility test was conducted to compare the resistance rates among Campylobacter spp. isolates from dogs (n = 50) raised under diverse conditions and humans (n = 50). More than 60% of Campylobacter (C.) jejuni from dogs and humans showed resistance to nalidixic acid, enrofloxacin and ciprofloxacin. C. jejuni isolates from humans showed higher resistance to tetracycline (83.3%) and ampicillin (91.3%) than those from dogs. None of the C. jejuni or Campylobacter coli isolates from humans or dogs were resistant to erythromycin. Overall, 85% of Campylobacter spp. isolates showed a multidrug resistant phenotype. Nucleotide sequencing analysis of the gryA gene showed that 100% of $NA^R/CIP^R$ C. jejuni isolates from dogs and humans had the Thr-$86^{th}$-Ile mutation, which is associated with fluoroquinolone resistance. flaA PCR restriction fragment length polymorphism (RFLP) typing to differentiate the isolates below the species level revealed 12 different clusters out of 73 strains. The human isolates belonged to eight different RFLP clusters, while five clusters contained dog and human isolates.

Unraveling Haplotype Diversity of the Apical Membrane Antigen-1 Gene in Plasmodium falciparum Populations in Thailand

  • Lumkul, Lalita;Sawaswong, Vorthon;Simpalipan, Phumin;Kaewthamasorn, Morakot;Harnyuttanakorn, Pongchai;Pattaradilokrat, Sittiporn
    • Parasites, Hosts and Diseases
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    • v.56 no.2
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    • pp.153-165
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    • 2018
  • Development of an effective vaccine is critically needed for the prevention of malaria. One of the key antigens for malaria vaccines is the apical membrane antigen 1 (AMA-1) of the human malaria parasite Plasmodium falciparum, the surface protein for erythrocyte invasion of the parasite. The gene encoding AMA-1 has been sequenced from populations of P. falciparum worldwide, but the haplotype diversity of the gene in P. falciparum populations in the Greater Mekong Subregion (GMS), including Thailand, remains to be characterized. In the present study, the AMA-1 gene was PCR amplified and sequenced from the genomic DNA of 65 P. falciparum isolates from 5 endemic areas in Thailand. The nearly full-length 1,848 nucleotide sequence of AMA-1 was subjected to molecular analyses, including nucleotide sequence diversity, haplotype diversity and deduced amino acid sequence diversity and neutrality tests. Phylogenetic analysis and pair-wise population differentiation ($F_{st}$ indices) were performed to infer the population structure. The analyses identified 60 single nucleotide polymorphic loci, predominately located in domain I of AMA-1. A total of 31 unique AMA-1 haplotypes were identified, which included 11 novel ones. The phylogenetic tree of the AMA-1 haplotypes revealed multiple clades of AMA-1, each of which contained parasites of multiple geographical origins, consistent with the $F_{st}$ indices indicating genetic homogeneity or gene flow among geographically distinct populations of P. falciparum in Thailand's borders with Myanmar, Laos and Cambodia. In summary, the study revealed novel haplotypes and population structure needed for the further advancement of AMA-1-based malaria vaccines in the GMS.

Authentication of Korean Panax ginseng from Chinease Panax ginseng and Panax quinquefolius by AFLP analysis

  • Kim Bo-Bae;Jeong Jae-Hun;Jung Su-Jin;Yun Doh-Won;Yoon Eui-Soo;Choi Yong-Eui
    • Journal of Plant Biotechnology
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    • v.7 no.2
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    • pp.81-86
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    • 2005
  • Panax ginseng is one of the most important medicinal plants in the world. The international trade of ginseng is increasing yearly. The disguise of Chinese and American ginseng into Korean ginseng became a problem in recent years in abroad and Korea. An effective method to authenticate the Korean Panax ginseng from others at a DNA level is necessary for the healthy development of the ginseng market. Amplified fragment length polymorphism (AFLP) analysis was applied to develop a method for the identification of Korean ginseng between Chinese ginseng and American ginseng. It is very difficult to detect the different polymorphic bands among Korean field cultivated ginseng, and between field and wild-cultivated ginseng. The genetic distance coefficient by AFLP analysis between field- and wild cultivated Korean ginseng was very low, 0.056. Whereas, polymorphic bands between Korean and Chinese wild-cultivated ginseng was significantly different. The genetic distance coefficient between wild-cultivated Korean and Chinese ginseng was 0.149. The genetic distance coefficients between the P. ginseng and P. quinquefolius were ranging from 0.626 to 0.666. These results support that the AFLP analysis could be applied to authenticate Korean P. ginseng from others Chinese P. ginseng and American ginseng (P. quinquefolius).

Morphological Characteristics and Genetic Relationship by RAPD Marker in Iris spp. (자생붓꽃의 형태적 특성 및 RAPD 마커에 의한 유연관계 분석)

  • Hong, Seong-Mi;Koh, Jae-Chul
    • Journal of Plant Biotechnology
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    • v.31 no.1
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    • pp.19-23
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    • 2004
  • This study was conducted to observe the morphological characteristics and to analyse genetic relationship using RAPD markers in 9 species of Iris. The period of flowering was shown very broad among 9 species of Iris, from April 22 to June 9. The flower stalk was fluctuated from 9.0cm (I. rossii) to 104.0cm (I. ensata). I. ensata has the largest flower size and followed by I. laevigata, I. tectorum, I. sanguinea, I. pseudacorus, I. lactea, I. rossii, I. minutoaurea, I. odaesanensis. Among the 108 amplified bands by PCR with 8 random primers, 107 showed polymorphism and only one showed monomorphism among 9 species of Iris. The average dissimilarity coefficient among 9 species of Iris was 0.252. The range of dissimilarity coefficient was shown as from 0.095 to 0.609. Nine species were divided into three groups. The first group was I. tectorum, and the second group was I. lactea, I. laevigata, I. pseudacorus, I. odaesanensis. I. minutoaurea, I. rossii, I. sanguinea, I. ensata were belong to group three.