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Isolation and Characterization of DNA Damaging Agent Sensitivity of rqh1 mutant from Schizosaccharomyce pombe (분열형 효모인 Schizosaccharomyces pombe 로부터 rqh1 돌연변이의 DNA damaging agent sensitivity를 보상하는 유전자의 특성 연구)

  • Lee, In-Hye;Choi, In-Soon
    • Journal of Life Science
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    • v.17 no.1 s.81
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    • pp.39-44
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    • 2007
  • The Rqh1 gene is essential for vegetative growth in fission Yeast. The rqh1 mutant showed that sensitivity of DNA damaging agent, a wild range of phenotype including abnormal gene expression and cell elongation. This result showed that the rqhl-overexpression cell was sensitivity to DNA damaging agent like rqhl mutant. When Rqh1 have an over-expression by $nmt1^+$ promoter of pREP vector, rqh1 mutant DNA damaging agent sensitivity could be compensated. We isolated two strong mutant containing complementation gene, rqh156 and rqh172, respectively. This result observed that the DNA damaging agent sensitivity of rqhl mutant was complemented by the expression of rqh156 and rqh172. They induced mRNA expression in a dose-dependent manner HU, MMS and UV. The HU sensitivity of the rqhl was complemented by the expression of rqh156 and rqh172. The mRNA expression of rqh156 decreased on HU dose dependent but the mRNA expression of rqh172 did not decrease on HU dose dependent. The MMS and W sensitivity of the rqhl was complemented by the expression of rqh156 and rqh172. These results indicate that the isolated rqhl gene may play an important role in DNA metabolism.

IscR Modulates Catalase A (KatA) Activity, Peroxide Resistance, and Full Virulence of Pseudomonas aeruginosa PA14

  • Kim, Seol-Hee;Lee, Bo-Young;Lau, Gee W.;Cho, You-Hee
    • Journal of Microbiology and Biotechnology
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    • v.19 no.12
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    • pp.1520-1526
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    • 2009
  • We have identified the iscR (PA3815) gene encoding an iron-sulfur cluster assembly regulator homolog as one of the genes required for peroxide resistance in Pseudomonas aeruginosa PA14. Here, we present the phenotypic characterization of an iscR deletion mutant in terms of KatA expression, stress responses, and virulence. The iscR null mutant exhibited reduced KatA activity at the posttranslational level, hypersensitivity to hydrogen peroxide, and virulence-attenuation in Drosophila melanogaster and mouse peritonitis models. These phenotypes were fully restored by multicopy-based expression of katA. These results suggest that the requirement of IscR in P. aeruginosa is related to the proper activity of KatA, which is crucial for peroxide resistance and full virulence of this bacterium.

An Analysis of Pronunciation Errors in Word-initial Onglides in English and a Suggestion of Teaching Method (어두에 나타나는 상향 이중모음의 오류분석 및 지도방안 연구)

  • Choi, Ju-Young;Park, Han-Sang
    • Proceedings of the KSPS conference
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    • 2007.05a
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    • pp.183-186
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    • 2007
  • This study analyzes Korean high school students' pronunciation errors in word-initial onglides in English. For this study, 24 Korean high school students read 34 English words including glide-vowel sequences in word-initial positions and vowel-initial words in a frame sentence. The results showed 2 different error types: glide deletion and vowel distortion. After the analysis of the first recording, the subjects were taught how to pronounce glide-vowel sequences properly in a 60-minute class. Comparison of the analyses of the first and second recordings showed that the subjects improved on the pronunciation of glide-vowel sequences. After the training, the pronunciation errors of diphthongs unique to English, [$j_I$], decreased substantially. However, most subjects still had difficulties in pronouncing [$w{\mho}$], [wu], and [wo]. There was no significant correlation between English course grade and error reduction.

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vfr, A Global Regulatory Gene, is Required for Pyrrolnitrin but not for Phenazine-1-carboxylic Acid Biosynthesis in Pseudomonas chlororaphis G05

  • Wu, Xia;Chi, Xiaoyan;Wang, Yanhua;Zhang, Kailu;Kai, Le;He, Qiuning;Tang, Jinxiu;Wang, Kewen;Sun, Longshuo;Hao, Xiuying;Xie, Weihai;Ge, Yihe
    • The Plant Pathology Journal
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    • v.35 no.4
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    • pp.351-361
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    • 2019
  • In our previous study, pyrrolnitrin produced in Pseudomonas chlororaphis G05 plays more critical role in suppression of mycelial growth of some fungal pathogens that cause plant diseases in agriculture. Although some regulators for pyrrolnitrin biosynthesis were identified, the pyrrolnitrin regulation pathway was not fully constructed. During our screening novel regulator candidates, we obtained a white conjugant G05W02 while transposon mutagenesis was carried out between a fusion mutant $G05{\Delta}phz{\Delta}prn::lacZ$ and E. coli S17-1 (pUT/mini-Tn5Kan). By cloning and sequencing of the transposon-flanking DNA fragment, we found that a vfr gene in the conjugant G05W02 was disrupted with mini-Tn5Kan. In one other previous study on P. fluorescens, however, it was reported that the deletion of the vfr caused increased production of pyrrolnitrin and other antifungal metabolites. To confirm its regulatory function, we constructed the vfr-knockout mutant $G05{\Delta}vfr$ and $G05{\Delta}phz{\Delta}prn::lacZ{\Delta}vfr$. By quantifying ${\beta}-galactosidase$ activities, we found that deletion of the vfr decreased the prn operon expression dramatically. Meanwhile, by quantifying pyrrolnitrin production in the mutant $G05{\Delta}vfr$, we found that deficiency of the Vfr caused decreased pyrrolnitrin production. However, production of phenazine-1-carboxylic acid was same to that in the wild-type strain G05. Taken together, Vfr is required for pyrrolnitrin but not for phenazine-1-carboxylic acid biosynthesis in P. chlororaphis G05.

Hypersensitivity of Somatic Mutations and Mitotic Recombinations Induced by Heterocyclic amines and Aflatoxin $B_1$ in Transgenic Drosophila (형질전환 초파리에서 Heterocyclic Amines와 Aflatoxin $B_1$에 의한 체세포 돌연변이 유발의 고감수성에 관한 연구)

  • 최영현;유미애;이원호
    • Korean journal of applied entomology
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    • v.35 no.4
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    • pp.315-320
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    • 1996
  • The effects of 2-arnino-3-methyIimidazo[4,5-fq]u inoline (IQ), 2-amino-6dimethyl-dipyrido[l,2-a;3',2'-d] imidazole (Glu-P-1) and aflatoxin B1 (AFBI) on the mitotic recombinations and somatic chromosome mutations were investigated using the transgenic Drosophila bearing a chimeric gene consisting of a promoter region of Drosophila actin 5C gene and rat DNA polymerase $. For investigating mitotic recombinations and the somatic chromosome mutations, the heterozygous (mwhl+) strain possessing or lacking transgene pol P was used. The spontaneous frequency of small mwh spots, due to deletion or nondisjunction etc., in the non-transgenic w strain and the transgenic plpol $1-130 strain was 0.351 and 0.606, respectively. The spontaneous frequency (0.063) of large mwh spots, arising mostly from somatic recombination between the centromere and the locus mwh, in the transgenic plpol $1-130 strain, was about three times higher than that (0.021) of the non-transgenic w strain. The mutant clone frequencies of two types induced by two heterocyclic mines (IQ and Glu-P-1) and AFBl in the transformant pbol PI-130 were two or three times higher than those in the host strain w. These mean that rat DNA polymerase P participates at least in the somatic chromosome mutations and mitotic recombination processes. And the present results suggest that the transgenic Drosophl!~ used in this study can be used as a hypersensitive, in vivo short-term assaying system for various environmental mutagens.

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Analysis of polymorphic regions of Plasmodium vivax Duffy binding protein of Korean isolates

  • Kho, Weon-Gyu;Chung, Joon-Yong;Sim, Eun-Jeong;Kim, Dong-Wook;Chung, Woo-Chul
    • Parasites, Hosts and Diseases
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    • v.39 no.2
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    • pp.143-150
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    • 2001
  • The present study was designed to investigate polymorphism in Duffy binding protein (DBP) gene of Plasmodium vivax isolates of Korea. Thirty samples were obtained from P. vivax patients in Yonchon-gun, Kyonggi-do in 1998. The PCR products of the samples were subjected to sequencing and hybridization analyses of the regions II and IV of P. vivax DBP gene. Two genotypes, SK-1 and SK-2, were identified on the basis of amino acid substitution and deletion. The genotype of 10 isolates was SK-1 and that of 20 isolates was SK-2. Most of the predicted amino acids in the region ll of DBP gene were conserved between the Korean isolates and Belem strain except for 4-5 amino acid substitutions. In the region W of DBP, a 6-bp insert that was shown in the Sal-1 allele type was found in SK-1, and a 27-bp insert that was shown in the Papua New Guinea allele type was found in SK-2. In conclusion, the present findings suggest that two genotypes of P. vivax coexist in the endemic area of Korea.

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Genetic Diversity and Population Structure in Native Chicken Populations from Myanmar, Thailand and Laos by Using 102 Indels Markers

  • Maw, A.A.;Kawabe, Kotaro;Shimogiri, T.;Rerkamnuaychoke, W.;Kawamoto, Y.;Masuda, S.;Okamoto, S.
    • Asian-Australasian Journal of Animal Sciences
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    • v.28 no.1
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    • pp.14-19
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    • 2015
  • The genetic diversity of native chicken populations from Myanmar, Thailand, and Laos was examined by using 102 insertion and/or deletion (indels) markers. Most of the indels loci were polymorphic (71% to 96%), and the genetic variability was similar in all populations. The average observed heterozygosities ($H_O$) and expected heterozygosities ($H_E$) ranged from 0.205 to 0.263 and 0.239 to 0.381, respectively. The coefficients of genetic differentiation (Gst) for all cumulated populations was 0.125, and the Thai native chickens showed higher Gst (0.088) than Myanmar (0.041) and Laotian (0.024) populations. The pairwise Fst distances ranged from 0.144 to 0.308 among populations. A neighbor-joining (NJ) tree, using Nei's genetic distance, revealed that Thai and Laotian native chicken populations were genetically close, while Myanmar native chickens were distant from the others. The native chickens from these three countries were thought to be descended from three different origins (K = 3) from STRUCTURE analysis. Genetic admixture was observed in Thai and Laotian native chickens, while admixture was absent in Myanmar native chickens.

Development of Movable Detection System for Efficiency Measurement in 3-PM Liquid Scintillation Counting

  • Hwang, H-Y;Kwak, S.I;Cho, Y.H;Byun, J.I;Lee, H.Y;Seo, J.S;Kwak, J.Y;Lee, J.M;Lee, K.B;Park, T.S;Chung, K.H;Lee, C.W
    • Nuclear Engineering and Technology
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    • v.35 no.2
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    • pp.165-170
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    • 2003
  • We developed an improved 3-PM liquid scintillation counting (3-PM LSC) method in which three detectors can be displaced to back and forth directions, and a data acquisition system being able to provide the values for all parameters required for the method. The detectors are entirely located in a 20-mm lead chamber of an inner diameter of 500 mm. A saw-toothed gear ties up all detectors so as to move them uniformly, up to 50 mm with unit of 1 mm. The data acquisition system was designed in an integrated circuit to perform the necessary works such as fast amplification, discrimination, coincidence and logic analysis. It generates values of nine parameters among twelve's generated in the 3-PH LSC method. The dead time of each counting channel is of extending type, valving from 10 to 100 $mutextrm{s}$. We measured the TDCR values with an unquenched liquid scintillation source 1"C by displacing the detectors with a step of 2.5 mm away from counting vial. Their values were derived on the range from 0.9 to 0.6. The extent is three times wider than those regions observed by applying the defocalization technique.ique.

The Effect of Dietary Docosahexaenoic Acid Enrichment on the Expression of Porcine Hepatic Genes

  • Chang, W.C.;Chen, C.H.;Cheng, W.T.K.;Ding, S.T.
    • Asian-Australasian Journal of Animal Sciences
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    • v.20 no.5
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    • pp.768-774
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    • 2007
  • To study the effect of dietary docosahexaenoic acid (DHA) enrichment on the expression of hepatic genes in pigs, weaned, crossbred pigs (30 d old) were fed diets supplemented with either 2% tallow or DHA oil for 18 d. Hepatic mRNA was extracted. Suppression subtractive hybridization was used to explore the hepatic genes that were specifically regulated by dietary DHA enrichment. After subtraction, we observed 288 cDNA fragments differentially expressed in livers from pigs fed either 2% DHA oil or 2% tallow for 18 d. After differential screening, 7 genes were found to be differentially expressed. Serum amyloid A protein 2 (SAA2) was further investigated because of its role in lipid metabolism. Northern analysis indicated that hepatic SAA2 was upregulated by dietary DHA enrichment (p<0.05). In a second experiment, feeding 10% DHA oil for 2d significantly increased the expression of SAA2 (compared to the 10% tallow group; p<0.05). The porcine SAA2 full length cDNA sequence was cloned and the sequence was compared to the human and mouse sequences. The homology of the SAA2 amino acid sequence between pig and human was 73% and between pig and mouse was 62%. There was a considerable difference in SAA2 sequences among these species. Of particular note was a deletion of 8 amino acids, in the pig compared to the human. This fragment is a specific characteristic for the SAA subtype that involved in acute inflammation reaction. Similar to human and mouse, porcine SAA2 was highly expressed in the liver of pigs. It was not detectable in the skeletal muscle, heart muscle, spleen, kidney, lung, and adipose tissue. These data suggest that SAA2 may be involved in mediation of the function of dietary DHA in the liver of the pig, however, the mechanism is not yet clear.