• Title/Summary/Keyword: pyrH

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Screening of Growth- or Development-related Genes by Using Genomic Library with Inducible Promoter in Aspergillus nidulans

  • Lee Bang-Yong;Han Sang-Yong;Choi Han Gil;Kim Jee Hyun;Han Kap-Hoon;Han Dong-Min
    • Journal of Microbiology
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    • v.43 no.6
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    • pp.523-528
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    • 2005
  • Using the genomic library constructed at the downstream of the niiA promoter, which induces the over-expression of an inserted DNA fragment, we have attempted to screen the genes affecting growth or development by over-expression. The wild-type strain was transformed using the AMA-niiA(p) library and cultured on 1.2 M sorbitol media, in which asexual sporulation is induced, but sexual development is repressed. Over 100,000 strains transformed to $pyrG^+$ were analyzed with regard to any changes in phenotype. Consequently, seven strains were isolated for further analyses. These strains were designated NOT [niiA(p) over-expression transformants] stains. Four of the strains were of the inducible type, and the remaining strains were of the multi-copy suppression type. Two of the inducible-type strains, NOT 1 and NOT40, harbored genes which had been inserted in reverse direction, suggesting that the mutant phenotypes had been derived from an excess amount of anti-sense mRNA. Domain analyses of the deduced polypeptides from the DNA fragments rescued from the transformants revealed that NOT1, NOT40 and NOT6 harbored a LisH motif, a forkhead domain, and a $Zn(II)_2Cys_6$ binuclear zinc cluster, respectively.

Metabolic Adjustments of Lactate Dehydrogenase Isozymes to the Environmental Temperature in Bluegill (Lepomis macrochirus) (환경온도에 대한 파랑볼우럭(Lepomis macrochirus) 젖산탈수소효소 동위효소들의 대사조절)

  • Ku, Bora;Cho, Sung Kyu;Yum, Jung Joo
    • Journal of Life Science
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    • v.26 no.10
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    • pp.1105-1112
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    • 2016
  • The aim of this study was to examine the metabolic adjustment of lactate dehydrogenase (EC 1.1.1.27, LDH) isozymes to the environmental temperature in bluegill (Lepomis macrochirus). This study included three groups of bluegill collected in April (group Ⅰ), May (group Ⅱ), and September (group Ⅲ). The LDH activities of skeletal muscle, heart, and brain tissues were higher in group Ⅲ than in groups Ⅰ and Ⅱ. The citrate synthase (EC 4.1.3.7, CS) activity was higher in skeletal muscle but lower in heart and brain tissues of group Ⅱ as compared to group Ⅰ. In contrast, the CS activity was lower in skeletal muscle and higher in heart and brain tissues in group Ⅲ than in group Ⅱ. Furthermore, the LDH/CS activity ratio was higher in the skeletal muscle and brain in group Ⅲ than in groups Ⅰ and Ⅱ. Accordingly, anaerobic metabolism was increased in group Ⅲ. LDH A4, A2B2, and B4 isozymes were expressed in skeletal muscle, heart, liver, and brain tissues. The LDH C hybrid was detected in brain tissue. The LDH A4 isozyme was successfully purified by affinity chromatography. The molecular weight of the purified LDH A4 isozyme was 136 kDa and its optimal pH for enzymatic activity was 8.0. The KmPYR values of LDH in skeletal muscle were 0.161-0.227 mM using pyruvate as a substrate. These kinetic properties of LDH in skeletal muscle are consistent with the fact that bluegill is a cold-adapted species. These results may be useful for predicting the habitat use of this fish.

Herbicidal activity of a new benzenesulfonylurea compound, KSC-13906 (신규 benzenesulfonylurea 계 화합물 KSC-13906의 제초활성)

  • Hwang, I.T.;Choi, J.S.;Ko, Y.K.;Choi, Y.S.;Kim, M.H.;Kim, D.W.;Cho, K.Y.
    • The Korean Journal of Pesticide Science
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    • v.3 no.1
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    • pp.1-12
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    • 1999
  • Herbicidal activities of newly synthesized experimental compound, KSC-13906 [Erythro N-{(4,6-dimethoxy pyrimidin-2-yl)aminocarbgnyl}-2-(2-fluoro-1- hydroxy-n-propyl)benzenesulfonamide, US Patent 5,461,025], were investigated in greenhouse and field for development as a paddy herbicide. KSC-13906 exihibited excellent weeding effects and selectivity to rice under submerged paddy condition in geenhouse. At the application rate of $2.5{\sim}5$ g ai/ha treated on surface of water, KSC-13906 effectively controlled three annual weeds such as Echinochloa crus-galli var. oryzicola. Scirpus juncoides, and Monochoria vaginalis as well as five perennial weeds such as Cyperus serotinus, Sagittaria pygmaea, Eleocharis kuroguwai, Potamogeton distinctus, and S. trifolia. However, there was no effect on Aneilema keisak and Lindernia procumbens. Especially, KSC-13906 effectively controlled Echinochloa crus-galli var. oryzicola at growth stages ranging from pre-emergence to 3-leaf stage. Antagonistic interaction was found between pyrazosulfuron-ethyl and mefenacet, but not between KSC-13906 ana mefenacet. KSC-13906 had high selectivity to transplanted rice, but caused phytotoxic effect on direct-seeded rice when treated before 3-leaf stage. Herbicidal activity of KSC-13906 on the Echinochloa crus-galli var. oryzicola was 7-folds greater than that of pyrazosulfuron-ethyl in terms of relative dose ratio of the herbicides. These results suggest that KSC-13906 possesses selective herbicidal potency in rice.

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Characterization of Lactate Dehydrogenase in Acanthogobius hasta (풀망둑(Acanthogobius hasta) 젖산탈수소효소의 특성)

  • Yum, Jung-Joo
    • Journal of Life Science
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    • v.18 no.2
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    • pp.264-272
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    • 2008
  • The lactate dehydrogenase (EC 1.1.1.27, LDH) isozymes in tissues from Acanthogobius hasta were characterized by biochemical, immunochemical and kinetic methods. The activities of LDH in skeletal muscle and eye tissues were 65.30 and 53.25 units, but LDH activities in heart and liver tissues were very low. LDH/CS (EC 4.1.3.7, citrate synthase) in skeletal muscle was the highest as 22.29. Specific activities of LDH in brain, eye and skeletal muscle were 56.45, 38.04 and 11.0 units/mg, respectively. The LDH isozymes in tissues were separated by polyacrylamide gel electrophoresis after immunoprecipitation with antiserum against $A_4,\;B_4$ eye-specific $C_4$ and liver-specific $C_4$. LDH $AC_4$ isozymes were detected predominantly in skeletal muscle, brain and eye tissues, and $B_4$ isozyme was detected in heart. Anodal eye-specific $C_4$ and cathodal liver-specific $C_4$ were coexpressed in A. hasta. The eye-specific $C_4$ isozyme showed higher activity in eye tissue, but liver-specific $C_4$ isozyme showed lower activity in liver. As a result, one part of molecular structures in $A_4\;and\;C_4,\;A_4\;and\;B_4$, and eye-specific $C_4$ and liver-specific $C_4$ were similar, but in $B_4\;and\;C_4$ were different with each other. Therefore the subunit A may be conservative in evolution, and the evolution of subunit B seems to be faster than that of subunit A. The LDH $A_4$ isozyme of skeletal muscle was purified in the fraction from elution with NAD+ containing buffer of affinity chromatography and eye-specific $C_4$ isozyme was eluted right after $A_4$, so the structure of eye-specific $C_4$ isozyme is similar to $A_4$. And LDH activity remained 35.22-43.47% as a result of the inhibition by pyruvate, the Michaelis-Menten constant values for pyruvate was 0.080-0.098 mM, and Vmax were 153.85 units, 35.09 units in skeletal muscle and eye, respectively. Also the $B_4$ isozyme was the thermo-stablest and $C_4$ was stabler than $A_4$ isozyme. The optimum pH of LDH was 6.5. The results mentioned above indicate that isozymes in tissues showed the properties between LDH $A_4\;and\;B_4$ isozyme as A. hasta was adapted to hypoxic conditions. Also LDH seems to function more effectively under anaerobic condition because LDH in skeletal muscle and eye tissues have high affinity for pyruvate.