• The Plant Pathology Journal
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• 제21권4호
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• pp.349-354
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• 2005

A virus isolate collected from infected paprika (Capsicum annuum var. grossum) was characterized as Potato virus Y (PVY) based on biological, serological, cytopathological, and molecular properties. In host range studies, the paprika isolate produced the mosaic symptom on some tobacco, tomato and pepper (Capsicum annuum). A new paprika isolate also infected potato cultivars which is different biological characteristic compared to the other popular potyvirus infecting paprika, Pepper mottle virus (PepMoV). Previously reported PVY strains, $PVY^o$ and $PVY^N$ did not infect pepper and typical PepMoV isolates did not infect potato. Distinctive inclusion patterns of the scroll, pinwheel, long laminated inclusions, and helper components in the cytoplasm of infected cells were also different to those observed by the typical PVY isolate infections. However, the paprika isolate reacted to the monoclonal antibody of $PVY^N$ strain with high absorbance readings. RT-PCR amplification, cloning, and sequencing of the 3' untranslated region and a part of coat protein gene also added additional evidence of the paprika isolate as the $PVY^N$-related isolate. Multiple alignments as well as cluster dendrograms of PVY-paprika isolate revealed close phylogenetic relationship to the $PVY^N$ subgroup. Altogether, these results suggest that a new PVY isolate infecting paprika contained distinct characteristics compared to the other previously described PVY strains with closer relationship to the $PVY^N$ strain.

• Biotechnology and Bioprocess Engineering:BBE
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• 제8권2호
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• pp.126-129
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• 2003

A relatively high homology between Escherichia coli prolidase and Alteromonas organophosphorous acid anhydrolase suggests that E. coli prolidase may have an activity to d egrade toxic organophosphorous compounds. To confirm this suggestion, we cloned and expressed a prolidase gene (pepQ) of E. coliBL2l. The recombinant E. coli prolidase that consisted of 443 amino acid residues exhibited activity and stereochemical selectivity against organopho sphorous compounds, although its activity was two to three orders of magnitude less than that of the other organophosphorous acid hydrolase isolated from Pseudomonas diminuta.

• The Plant Pathology Journal
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• 제24권1호
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• pp.93-96
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• 2008

In August 2006, a severe disease incidence showing mosaic and/or necrotic symptoms on two bell pepper varieties including red-colored 'Special' and yellow-colored 'Fiesta' was observed in a greenhouse located in Gwangyang, Jeonnam province, Korea. To identify causal viruses, total RNAs were extracted from 11 fruit samples with and without symptoms. Specific oligonucleotide primers for Cucumber mosaic virus (CMV), Pepper mottle virus (PepMoV), Tomato spotted wilt virus (TSWV) and Pepper mild mottle virus (PMMoV) were designed based on the sequences available on GenBank. Database comparisons of the deduced amino acid sequences of each sequence produced 100% and 98% matches with nucleocapsid protein gene of TSWV (Acc. No. ABE11605) and coat protein gene of CMV (Acc. No. DQ018289), respectively, suggesting that the symptoms on bell pepper fruits might be caused by the infection of CMV and TSWV. To our knowledge this is the first report of necrotic as well as mosaic virus disease on bell pepper fruits by the infection of CMV and TSWV in Jeonnam province, Korea.

• 한국한의학연구원논문집
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• 제8권1호
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• pp.109-126
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• 2002

The herbal extract (YMT_02) is a modified herbal extracts from Yukmijihwangtang (YMJ) to promote memory-enhancing. The YMJ extracts has been widely used as an anti-aging herbal medicine for hundred years in Asian countries. The purpose of this study is to; 1) quantitatively evaluate the memory-enhancing effect of YMT_02 by hehavior task, 2) identify candidate genes responsible for enhancing memory by cDNA microarray and 3) assess the anti-oxidant effect of YMT_02 on PC12 cell. Memory retention abilities are addressed by passive avoidance task with Sprague-Dawley (SD) male rat. Before the training session, the rats are subdivided into four groups and administrated with YMT_02, Ginkgo biloba, Soya lecithin and normal saline for 10 days. The retention test was performed. 24 hours after the training session. The retention time of the YMT_02 group was significantly (p<0.05) delayed $({\sim}100%)$, whereas Ginkgo biloba and Soya lecithin treatment delayed 20% and 10% respectively. The hippocampi of YMT_02 and control group were dissected and mRNA was further purified. After synthesizing cDNA using oligo-dT primer, the cDNA were applied and mRNA was further purified. After synthesizing cDNA using oligo-dT primer, the cDNA were applied to Incyte rat GEMTM 2 cDNA microarray. The microarray results show that prealbumin(transthyretin), phosphotidy lethanolamine N-methyltransferase, and PEP-19 are expressed abundantly in the YMT_02 treated group. Especially, PEP-19 is a neuron-specific protein, which inhibits apoptotic processes in neuronal cell. On the other hand, transcripts of RAB15, glutamate receptor subunit 2 and CDK 108 are abundant in control group. Besides, neuronal genes involved in neuronal death or neurodegeneration such as neuronal-pentraxin and spectrin are abundantly expressed in control group. Additionally, the YMT_02 shows an anti oxidative effect in the PC12 cell. The list of differentially expressed genes may implicate further insight on the action and mechanism behind the memory-enhancing effect of herbal extracts YMT_02, for example, anti-apoptotic, anti-oxidative, and neuroprotective effects.

• Animal Bioscience
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• 제35권1호
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• pp.64-74
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• 2022

Objective: This study was conducted to investigate the effects of dietary multi-strain probiotic (MSP) (Bacillus coagulans Unique IS2 + Bacillus subtillis UBBS14 + Saccharomyces boulardii Unique 28) on performance, gut morphology and expression of nutrient transporter related genes in broiler chickens. Methods: A total of 256 (4×8×8) day-old CARIBRO Vishal commercial broiler chicks of uniform body weight were randomly distributed into four treatments with 8 replicates each and having eight chicks in each replicate. Four dietary treatments were T₁ (negative control-basal diet), T₂ (positive control-antibiotic bacitracin methylene disalicylate at 20 mg/kg diet), T₃ (MSP at 10⁷ colony-forming unit [CFU]/g feed), and T₄ (MSP at 10⁸ CFU/g feed). Results: During 3 to 6 weeks and 0 to 6 weeks, the body weight gain increased significantly (p<0.05) in T₃ and T₄ groups. The feed intake significantly (p<0.05) reduced from T₁ to T₃ during 0 to 3 weeks and the feed conversion ratio also significantly (p<0.05) improved in T₃ and T₄ during 0 to 6 weeks. The humoral and cell mediated immune response and the weight of immune organs were also significantly (p<0.05) improved in T₃ and T₄. However, significant (p<0.05) dietary effects were observed on intestinal histo-morphometry of ileum in T₃ followed by T₄ and T₂. At 14 d post hatch, the relative gene expression of glucose transporter (GLUT5), sodium-dependent glucose transporter (SGLT₁) and peptide transporter (PepT₁) showed a significant (p<0.05) up-regulating pattern in T₂, T₃, and T₄. Whereas, at 21 d post hatch, the gene expression of SGLT₁ and PepT₁ was significantly (p<0.05) downregulated in MSP supplemented treatments T₃ and T₄. Conclusion: The supplementation of MSP at 10⁷ CFU/g diet showed significant effects with improved performance, immune response, gut morphology and expression of nutrient transporter genes. Thus, the MSP could be a suitable alternative to antibiotic growth promoters in chicken diets.

• Journal of Genetic Medicine
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• 제2권2호
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• pp.53-57
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• 1998

Spinal muscular atrophy (SMA) type I is a common severe autosomal recessive inherited neuromuscular disorder that has been mapped to chromosome 5q11.2-13.3. The survival motor neuron (SMN) gene, a candidate gene, is known to be deleted in 96% of patients with SMA type I. Presently, PCR and single strand conformation polymorphism (PCR-SSCP) analyses have been made possible for application to both archival slides and paraffin-embedded tissues. Archival materials represent valuable DNA resources for genetic diagnosis. We applied these methods for the identification of SMN gene of SMA type I in archival specimens for the prenatal diagnosis. In this study, we performed the prenatal diagnosis with chorionic villus sampling (CVS) cells on two women who had experienced neonatal death of SMA type I. DNA extraction was done from archival slide and tissue materials and PEP-PCR was performed using CVS cells. In order to identify common deletion region of SMN and neuronal apoptosis-inhibitory protein (NAIP) genes, cold PCR-SSCP and PCR-restriction site assay were carried out. Case 1 had deletions of the exons 7 and 8, and case 2 had exon 7 only on the telomeric SMN gene. Both cases were found to be normal on NAIP gene. These results were the same for both CVS and archival biopsied specimens. In both cases, the fetuses were, therefore, predicted to be at very high risk of being affected and the pregnancy were terminated. These data clearly demonstrate that archival slide and paraffin-embedded tissues can be a valuable source of DNA when the prenatal genetic diagnosis is needed in case any source for genetic analysis is not readily available due to previous death of the fetus or neonate.

• Journal of Microbiology and Biotechnology
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• 제18권4호
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• pp.746-753
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• 2008

The hprK gene encoding bifunctional HPrK/P (kinase/phosphorylase) was cloned from L. mesenteroides SY1, a strain isolated from kimchi. hprK was transcribed as a monocistronic gene. His-tagged HPrH16A and HPrK/P were produced in E. coli BL21 (DE3) using pET26b(+) and purified. HPrK/P phosphorylation assay with purified proteins showed that the kinase activity of HPrK/P increased at slightly acidic pHs. Divalent cations such as $Mg^{2+}$ and $Mn^{2+}$ and glycolytic intermediates such as fructose-1, 6-bisphosphate (FBP) and phosphoenolpyruvate (PEP) increased the kinase activity of HPrK/P, but inorganic phosphate strongly inhibited it. Kinetic studies for the kinase activity of HPrK/P showed that the apparent $K_m$ values were 0.18 and $14.57{\mu}M$ for ATP and HPr, respectively. The $K_m$ value for the phosphorylase activity of HPrK/P was $14.16{\mu}M$ for P-Ser-HPr (HPr phosphorylated at the serine residue).

• 동의생리병리학회지
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• 제16권5호
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• pp.1025-1034
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• 2002

The herbal extract (YMT_02) is a modified herbal extracts from Yukmijihwang-tang (YMJ) to promote memory-enhancing. The YMJ extracts has been widely used as an anti-aging herbal medicine for hundred years in Asian countries. The purpose of this study is to; 1) quantitatively evaluate the memory-enhancing effect of YMT_02 by behavior task, 2) identify candidate genes responsible for enhancing memory by cDNA microarray and 3) assess the anti-oxidant effect of YMT_02 on PC12 cell. Memory retention abilities are addressed by passive avoidance task with Sprague-Dawley (SD) male rat. Before the training session, the rats are subdivided into four groups and administrated with YMT_02, Ginkgo biloba, Soya lecithin and normal saline for 10 days. The retention test was performed. 24 hours after the training session. The retention time of the YMT_02 group was significantly (p<0.05) delayed (～100%), whereas Ginkgo biloba and Soya lecithin treatment delayed 20% and 10% respectively. The hippocampi of YMT_02 and control group were dissected and mANA was further purified. After synthesizing cDNA using oligo-dT primer, the cDNA were applied to Incyte rat GEMTM 2 cDNA microarray. The microarray results show that prealbumin(transthyretin), phosphotidylethanolamine N-methyltransferase, and PEP-19 are expressed abundantly in the YMT_02 treated group. Especially, PEP-19 is a neuron-specific protein, which inhibits apoptotic processes in neuronal cell. On the other hand, transcripts of RAB15, glutamate receptor subunit 2 and CDK108 are abundant in control group. Besides, neuronal genes involved in neuronal death or neurodegeneration such as neuronal-pentraxin and spectrin are abundantly expressed in control group. Additionally, the YMT_02 shows an anti oxidative effect in the PC12 cell. The list of differentially expressed genes may implicate further insight on the action and mechanism behind the memory-enhancing effect of herbal extracts YMT_02, for example, anti-apoptotic, anti-oxidative, and neuroprotective effects.

• 원예과학기술지
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• 제28권4호
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• pp.648-655
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• 2010

본 연구에서는 들잔디와 돌연변이체에서 Glyphosate 내성 관련 유전자인 EPSPS를 코딩하는 cDNA를 분리하여, 시퀸스 비교분석과 발현 양상 차이 등에 관하여 조사하였다. 5'/3' RACE를 통하여 밝혀진 EPSPS의 cDNA는 각각 1176bp의 open reading frame으로 이루어져 있으며 391개의 아미노산을 코딩하고 있었다. 이는 보고된 다른 EPSPS 유전자들과 높은 유사성을 가지고 있다. Genomic southern 결과 들잔디 내에 EPSPS 유전자는 단일copy로 존재하였다. Wild type과 제초제 내성 변이체의 EPSPS 유전자는 6개의 아미노산 시퀀스의 차이를 보였으며 기존에 보고된 EPSPS active site에서 시퀀스의 차이를 나타냈다. 한편 glyphosate의 독성기작의 하나인 EPSPS 효소활성 저해 작용을 알아본 결과, 내성 개체에서 높은(1.5배 이상) EPSPS 활성을 확인할 수 있었다. Northern 분석과 RT-PCR로 제초제 처리 시간에 따른 EPSPS의 발현량을 살펴본 결과, 제초제 처리 후 시간이 경과할수록 발현량이 증가하다가 7일 이후에는 감소하였고, wild type에 비해 glyphosate 내성 선발개체에서 전체적인 발현량이 높음을 알 수 있었다. 본 연구 결과 선발된 glyphosate 내성 돌연변이체는 높은 EPSPS 효소활성 증가 및 EPSPS active site의 아미노산 시퀀스 변화를 통해 원할하고 지속적인 방향족 아미노산의 합성이 가능한 것으로 보여졌다. 본 실험을 통해 얻어진 glyphosate 내성 잔디 돌연변이체와 방법은 앞으로 유용한 제초제 저항성 돌연변이 품종개발 연구에 활용될 수 있을 것으로 기대된다.

• Journal of Microbiology and Biotechnology
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• 제25권7호
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• pp.1070-1083
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• 2015

A gene (GT-SM3B) encoding a thermostable secreted oligoendopeptidase (GT-SM3B) was cloned from the thermophile Geobacillus thermoleovorans DSM 15325. GT-SM3B is 1,857 bp in length and encodes a single-domain protein of 618 amino acids with a 23-residue signal peptide having a calculated mass of 67.7 kDa after signal cleavage. The deduced amino acid sequence of GT-SM3B contains a conservative zinc metallopeptidase motif (His⁴⁰⁰-Glu⁴⁰¹-X-XHis⁴⁰⁴). The described oligopeptidase belongs to the M3B subfamily of metallopeptidases and displays the highest amino acid sequence identity (40.3%) to the oligopeptidase PepF_Ba from mesophilic Bacillus amyloliquefaciens 23-7A among the characterized oligopeptidases. Secretory production of GT-SM3B was used, exploiting successful oligopeptidase signal peptide recognition by Escherichia coli BL21 (DE3). The recombinant enzyme was purified from the culture fluid. Homodimerization of GT-SM3B was determined by SDS-PAGE. Both the homodimer and monomer were catalytically active within a pH range of 5.0–8.0, at pH 7.3 and 40℃, showing the K_m, V_max, and k_cat values for carbobenzoxy-Gly-Pro-Gly-Gly-Pro-Ala-OH peptidolysis to be 2.17 ± 0.04 × 10^-6 M, 2.65 ± 0.03 × 10^-3 µM/min, and 5.99 ± 0.07 s^-1, respectively. Peptidase remained stable at a broad pH range of 5.0–8.0. GT-SM3B was thermoactive, demonstrating 84% and 64% of maximum activity at 50℃ and 60℃, respectively. The recombinant oligopeptidase is one of the most thermostable M3B peptidase, retaining 71% residual activity after incubation at 60℃ for 1 h. GT-SM3B was shown to hydrolyze a collagenous peptide mixture derived from various types of collagen, but less preferentially than synthetic hexapeptide. This study is the first report on an extracellular thermostable metallo-oligopeptidase.