• Title/Summary/Keyword: Residues

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Cloning and Expression of a Alkaline Protease from Bacillus clausii I-52 (Bacillus clausii I-52로부터 alkaline protease 유전자의 클로닝 및 발현)

  • Joo, Han-Seung;Choi, Jang Won
    • Journal of agriculture & life science
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    • v.45 no.6
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    • pp.201-212
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    • 2011
  • The alkaline protease gene was cloned from a halo-tolerant alkalophilic Bacillus clausii I-52 isolated from the heavily polluted tidal mud flat of West Sea in Inchon Korea, which produced a strong extracellular alkaline protease (BCAP). Based on the full genome sequence of Bacillus subtilis, PCR primers were designed to allow for the amplification and cloning of the intact pro-BCAP gene including promoter region. The full-length gene consists of 1,143 bp and encodes 381 amino acids, which includes 29 residues of a putative signal peptide and an additional 77-amino-acid propeptide at its N-terminus. The mature BCAP deduced from the nucleotide sequence consists of 275 amino acids with a N-terminal amino acid of Ala, and a relative molecular weight and pI value was 27698.7 Da and 6.3, respectively. The amino acid sequence shares the highest similarity (99%) to the nattokinase precursor from B. subtilis and subtilisin E precursor from B. subtilis BSn5. The substrate specificity indicated that the recombinant BCAP could hydrolyze efficiently the synthetic substrate, N-Suc-Ala-Ala-Pro-Phe-pNA,and did not hydrolyze the substrates with basic amino acids at the P1 site. The recombinant BCAP was strongly inhibited by typical serine protease inhibitor, PMSF, indicating that BCAP is a member of the serine proteases.

Molecular cloning and expression pattern of Metallothionein Gene from the left-handed shell, Physa acuta (왼돌아물달팽이 (Physa acuta) 의 Metallothionein 유전자 클로닝 및 발현양상)

  • Jo, Yong-Hun;Baek, Moon-Ki;Kang, Se-Won;Lee, Jae-Bong;Byun, In-Seon;Choi, Sang-Haeng;Chae, Sung-Hwa;Kang, Jung-Ha;Han, Yeon-Soo;Park, Hong-Seog;Lee, Yong-Seok
    • The Korean Journal of Malacology
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    • v.25 no.3
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    • pp.223-230
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    • 2009
  • Metallothioneins (MTs) play a key role in metallic homeostasis and detoxification in most living organisms. In an attempt to study the biological functions and significance of MT in a snail, we cloned and partially characterized the MT gene from the left-handed snail, Physa acutawhich has been regarded as a potential biomonitering species for fresh water. The complete cDNA sequence of PaMT cDNA was identified from the expressed sequence tag (EST) sequencing project of Physa acuta. The coding region of 180 bp gives 60 amino acid residues including the initiation methionine and termination codon. Clustering and phylogenic analysis of PaMT with other MT amino acid sequences show that it has some identities to Helix pomatia (60%), Arianta arbustorum (58%), Perna viridis (49%), Mytilus edulis (49%), Bathymodiolus azoricus (49%), Bathymodiolus azoricus (48%) and Bathymodiolus sp. FD-2002 (48%). Time dependent induction for PaMT from P. acuta exposed with cadmium (50 ppb) indicated that PaMT was induced at 4-8 hr after exposure. It remains to further develop PaMTas a potential biomarker for water contamination in fresh water.

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Identification, sequence characterization and expression analysis of the arginine kinase gene in response to laminarin challenge from the Oriental land snail, Nesiohelix samarangae (동양달팽이(Nesiohelix samarangae)의 arginine kinase 유전자 분석 및 발현 패턴에 관한 연구)

  • Jeong, Ji Eun;Lee, Yong Seok
    • The Korean Journal of Malacology
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    • v.29 no.3
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    • pp.171-179
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    • 2013
  • Arginine kinase (ArK) is known to play an important role in most invertebrates the level of ATP by phosphorylation of phosphagens in cell and immuninty in living organisms. ArK has been identified in many kinds of organisms ranging from invertebrate to vertebrate. However, no ArK gene has been cloned and investigated from N. samarangae. This leads us to identify ArK cDNA (NsArK) from the expressed sequence tag (EST) sequencing of N. samarangae. Sequence analysis indicated that the coding region of 1,065 bp contains 355 amino acid residues. Molecular phylogenetic analysis shows that NsArK had very high similarities with mollusca and arthropoda. In an attempt to investigate a potential role of NsArK in the digestive gland of N. samarangae, expression patterns were analyzed. RT-PCR analsysis shows that NsArK mRNA is induced in the rane of 1.2 fold at 6 hr by laminarin when compared with the control. The immunnologial and physiological role of NsArK remains to be further investigated in N. samarangae.

Biomonitoring of Toxic Effects of Pesticides in Occupationally Exposed Individuals

  • Arshad, Muhammad;Siddiqa, Maryam;Rashid, Saddaf;Hashmi, Imran;Awan, Muhammad Ali;Ali, Muhammad Arif
    • Safety and Health at Work
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    • v.7 no.2
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    • pp.156-160
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    • 2016
  • Background: Workers in pesticide manufacturing industries are constantly exposed to pesticides. Genetic biomonitoring provides an early identification of potential cancer and genetic diseases in exposed populations. The objectives of this biomonitoring study were to assess DNA damage through comet assay in blood samples collected from industry workers and compare these results with those of classical analytical techniques used for complete blood count analysis. Methods: Samples from controls (n = 20) and exposed workers (n = 38) from an industrial area in Multan, Pakistan, were subjected to various tests. Malathion residues in blood samples were measured by gas chromatography. Results: The exposed workers who were employed in the pesticide manufacturing industry for a longer period (i.e., 13-25 years) had significantly higher DNA tail length ($7.04{\mu}m$) than the controls ($0.94{\mu}m$). Workers in the exposed group also had higher white blood cell and red blood cell counts, and lower levels of mean corpuscular hemoglobin (MCH), MCH concentration, and mean corpuscular volume in comparison with normal levels for these parameters. Malathion was not detected in the control group. However, in the exposed group, 72% of whole blood samples had malathion with a mean value of 0.14 mg/L (range 0.01-0.31 mg/L). Conclusion: We found a strong correlation ($R^2=0.91$) between DNA damage in terms of tail length and malathion concentration in blood. Intensive efforts and trainings are thus required to build awareness about safety practices and to change industrial workers' attitude to prevent harmful environmental and anthropogenic effects.

Quality Evaluation and Residual Pesticides of Lettuce during Growth after Transplanting (정식(定植) 후 양상추의 품질평가 및 잔류농약 분석)

  • Youn, Aye-Ree;Kim, Byeong-Sam;Kim, Sang-Hee;Kwon, Ki-Hyun;Cha, Hwan-Soo
    • Food Science and Preservation
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    • v.14 no.2
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    • pp.124-130
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    • 2007
  • Lettuce (Lactuca Sativa L.) is the most popular salad vegetable crop The purpose of this study was to analyze the quality of lettuce, and residual pesticides in this vegetable, during growth after transplanting. The eatable weight was 40% of the total weight at the commencement of transplanting, but had doubted at harvest after transplanting. As lettuce grew after transplanting, the pH, the sugar level, and the moisture content increased slightly, and the color tended towards yellow rather than greenThe SPDA chlorophyll measurement was 16.84 at 42 day post-ransplantation, and increased to 26.44 at 67 day after transplanting. The contents of vitamin C were 2.5mg/100g wet weight at 60 d after transplantation, but decreased to 2.2mg/100g wet weight 67 day post-transplantation. Mineral (Ca, Fe, K, Mg, and Na) levels rose after transplantation, and attained their highest levels 60 day post-transplantation. The lettuce crop was analyzed for residues of each of 48 pesticides nt 54 after transplantation, and the only such chemical detected was chlorthalonil(0.03 0.25mg/kg wet weight).

Decomposition of Rice Straw and Compost in an Acid Sulfate Soil under Aerobic and Anaerobic Conditions (특이산성토양(特異酸性土壤)에 있어서 볏짚 및 퇴비(堆肥)의 분해(分解)에 관(關)한 연구(硏究))

  • Lee, Sang Kyu;Yoo, Ick Dong;Parr, James F.
    • Korean Journal of Soil Science and Fertilizer
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    • v.8 no.4
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    • pp.171-176
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    • 1975
  • The rate and extent of decomposition of rice straw and compost in an acid sulfate soil amended with urea and lime and incubated under aerobic and anaerobic(flooded) conditions were investigated in the laboratory. Results are summarized as follows: 1. The rate of compost(alone) decomposition in a flooded soil was more than twice as high as all other treatments, which included rice straw+urea, rice straw+lime, rice straw (alone), and compost+lime. Lime appeared to suppress the decomposition of compost in a flooded soil but actually enhanced its decomposition under aerobic conditions. 2. Compost decomposition in both anaerobic and aerobic environments was characterized by single maximum peak rates of $CO_2$ evolution that were reached soon after the start of incubation. 3. Both urea and lime greatly increased the rate and extent of rice straw decomposition in the soil when incubated aerobically, although urea had a greater effect than did liming. Decomposition rates were characterized by the appearance of two maximum peak rates, a greater primary peak and a smaller secondary peak. 4. The percent decomposition of rice straw in soil incubated aerobically was approximately half (10.8%) that of compost(23.1%). However, percent decomposition of these substrates in soil amended with lime was essentially the same; i.e., rice straw+lime (29.4%) and compost+lime(31.6%). 5. There is a need to investigate the possible interaction between the addition of lime (pH) and supplemental nitrogen applied to acid sulfate soils and how this interaction might affect the decomposition of organic wastes and residues.

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Assessment on Nitrous oxide (N2O) Emissions of Korea Agricultural Soils in 2009 (2009년 우리나라 농경지 토양에서의 N2O 배출량 평가)

  • Jeong, Hyun-Cheol;Kim, Gun-Yeob;Lee, Deog-Bae;Shim, Kyo-Moon;Lee, Seul-Bi;Kang, Kee-Kyung
    • Korean Journal of Soil Science and Fertilizer
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    • v.44 no.6
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    • pp.1207-1213
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    • 2011
  • This study was conducted to assess $N_2O$ emissions in agricultural soils of Korea. According to 1996 and 2006 IPCC (Intergovernmental Panel on Climate Change) methodology, $N_2O$ emission was calculated the sum of direct emission ($N_2O_{DIRECT}$) and indirect emission ($N_2O_{INDIRECT}$). To calculate $N_2O$ emissions, emission factor was used default of IPCC and activity data was used the food, agricultural, forestry and fisheries statistical yearbook of MIFAFF (Ministry for Food, Agriculture, Forestry and Fisheries). It was emitted 8,608 $N_2O$ Mg resulted from direct emission by application of chemical fertilizer and animal manure, input in n-fixation crops and input of crop residues and emissions converted $N_2O$ into $CO_2$ equivalent was 2,668 $CO_2$-eq Gg. Indirect emission as $N_2O_{(G)}$ (atmospheric deposition of $NH_3$ and $NO_X$) and $N_2O_{(L)}$ (leaching and runoffs) were 4,567 and 6,013 $N_2O$ Mg and emissions converted $N_2O$ into $CO_2$ equivalent were 1,416 and 1,864 $CO_2$-eq Gg, respectively. Total $N_2O$ emission in Korea agricultural soil in 2009 was 5,948 $CO_2$-eq Gg.

Antimicrobial activity and characterization for defensin of synthetic oligopeptides derived from Bombus ignitus (호박벌 유래 디펜신 유전자의 분자적 특성분석 및 항균 활성)

  • Kang, Heui-Yun;Kim, In-Woo;Lee, Joon-Ha;Kwon, Young Nam;Yun, Eun-Young;Yoon, Hyung Joo;Kim, Seong-Ryul;Kim, Iksoo;Hwang, Jae-Sam
    • Journal of Sericultural and Entomological Science
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    • v.50 no.2
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    • pp.161-165
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    • 2012
  • Antimicrobial peptides of insects are found and reported as immune defence system against infectious agents. The peptides are produced by fat body cells and thrombocytoids, a blood cell type. Defensin is 38-45 amino acids long and consists of an ${\alpha}$-helix linked by a loop to an antiparallel ${\beta}$-sheet. Defensin from a bumblebee, Bombus ignitus, is known to comprise 52 amino acid residues. This peptide consists of two ${\alpha}$-helixes; ACAANCLSM and KTNFKDLWDKRF and one ${\beta}$-sheet; GGRCENGVCLCR. We carried out antibacterial activity test by radial diffusion assay against Staphylococcus aureus (Gram positive), Escherichia coli (Gram negative), Pseudomonas syringae (Gram negative), Candida albicans (fungi), MDRPA, MRSA, and VRE (antimicrobial resistant microbes) with synthetic oligopeptides from Peptron (Daejeon, Korea). The predicted curtailment fragment (GGRCEVCLCR-$NH_2$) for ${\beta}$-sheet had strong antibacterial activity when internal amino acids were removed. But, curtailment fragments (ACAANCLSM-$NH_2$ and TNFKDLWDKR-$NH_2$) of ${\alpha}$-helix were not showed antibacterial activity. These synthetic oligopeptides were showed the great activity against Gram positive and negative bacteria.

Novel reassortant 2.3.4.4B H5N6 highly pathogenic avian influenza viruses circulating among wild, domestic birds in Xinjiang, Northwest China

  • Zhang, Qian;Mei, Xindi;Zhang, Cheng;Li, Juan;Chang, Nana;Aji, Dilihuma;Shi, Weifeng;Bi, Yuhai;Ma, Zhenghai
    • Journal of Veterinary Science
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    • v.22 no.4
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    • pp.43.1-43.10
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    • 2021
  • Background: The H5 avian influenza viruses (AIVs) of clade 2.3.4.4 circulate in wild and domestic birds worldwide. In 2017, nine strains of H5N6 AIVs were isolated from aquatic poultry in Xinjiang, Northwest China. Objectives: This study aimed to analyze the origin, reassortment, and mutations of the AIV isolates. Methods: AIVs were isolated from oropharyngeal and cloacal swabs of poultry. Identification was accomplished by inoculating isolates into embryonated chicken eggs and performing hemagglutination tests and reverse transcription polymerase chain reaction (RT-PCR). The viral genomes were amplified with RT-PCR and then sequenced. The sequence alignment, phylogenetic, and molecular characteristic analyses were performed by using bioinformatic software. Results: Nine isolates originated from the same ancestor. The viral HA gene belonged to clade 2.3.4.4B, while the NA gene had a close phylogenetic relationship with the 2.3.4.4C H5N6 highly pathogenic avian influenza viruses (HPAIVs) isolated from shoveler ducks in Ningxia in 2015. The NP gene was grouped into an independent subcluster within the 2.3.4.4B H5N8 AIVs, and the remaining six genes all had close phylogenetic relationships with the 2.3.4.4B H5N8 HPAIVs isolated from the wild birds in China, Egypt, Uganda, Cameroon, and India in 2016-2017, Multiple basic amino acid residues associated with HPAIVs were located adjacent to the cleavage site of the HA protein. The nine isolates comprised reassortant 2.3.4.4B HPAIVs originating from 2.3.4.4B H5N8 and 2.3.4.4C H5N6 viruses in wild birds. Conclusions: These results suggest that the Northern Tianshan Mountain wetlands in Xinjiang may have a key role in AIVs disseminating from Central China to the Eurasian continent and East African.

Immunostimulatory activity and structural characteristics of neutral polysaccharides isolated from ginseng leaves fermented by Cordyceps sinensis (동충하초균으로 발효한 인삼잎에서 분리한 중성다당의 면역활성 및 구조적 특성)

  • Cha, Ha Young;Park, Hye-Ryung;Shin, Kwang-Soon
    • Korean Journal of Food Science and Technology
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    • v.53 no.5
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    • pp.570-577
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    • 2021
  • This study elucidated the biological activities and structural characteristics of polysaccharides isolated from ginseng leaves fermented using Cordyceps sinensis (GLF). GLF comprised at least 18 glycosyl linkages, including 4-linked glucose residues (24.0%). To characterize the neutral polysaccharides in GLF, it was further fractionated by anion exchange chromatography, and the unabsorbed fraction (GLF1) was isolated. Peritoneal macrophages stimulated with GLF1 produced various cytokines in a dose-dependent manner. The properties and activities of the four subfractions (PHI, PHIA1-PHIA3) obtained after sequential enzymatic digestion were examined. PHI and PHIA3 primarily comprised glucose, whereas PHI exhibited an iodine-color reaction. Furthermore, the PHIA1-3 fractions indicated that cytokine production was completely inhibited. These results suggest at the immune activities of GLF1 may be due to the α-(1→4)-glucan branched at the C(O)6 position, which was produced by C. sinensis.