• Title/Summary/Keyword: Viral disease

Search Result 951, Processing Time 0.022 seconds

Expression of SARS-3CL Protease in a Cell-Free Protein Synthesis System (무세포 단백질 합성법을 이용한 활성형 SARS-3CL protease의 발현)

  • Park, Sun-Joo;Kim, Yong-Tae
    • Journal of Life Science
    • /
    • v.22 no.4
    • /
    • pp.552-558
    • /
    • 2012
  • Severe acute respiratory syndrome (SARS) is a severe respiratory infectious disease caused by a novel human coronavirus, SARS-CoV. The 3CL protease is a key enzyme in the proteolytic processing of replicase polyprotein precursors, pp1a and pp1ab, which mediate all the functions required for viral genomic replication and transcription. Therefore, this enzyme is a target for the development of chemotherapeutic agents against SARS. A large quantity of active SARS-3CL protease is required for development of anti-SARS agents. Here we have constructed overexpression vector for the production of the SARS-3CL protease. The gene encoding SARS-3CL protease was amplified using polymerase chain reaction and cloned into the pET29a expression vector, resulting in pET29a/SARS-3CLP. Recombinant SARS-3CL protease was successfully synthesized by the dialysis mode of the cell-free protein expression system, and purified by three-step fast protein liquid chromatography using HighQ and MonoP column chromatographies and Sephacryl S-300 gel filtration. In addition, the produced SARS-3CL protease was found to be an active mature form. This study provides efficient methods not only for the development of anti-SARS materials from natural sources, but also for the study of basic properties of the SARS-3CL protease.

Occurrence of Stone Fruit Viruses on Peach Trees (Prunus persica L. Batsch) in Korea (국내에서 발생하는 복숭아 바이러스병)

  • Cho, In Sook;Cho, Jeom Doeg;Choi, Seung Kook;Choi, Gug Seoun
    • Research in Plant Disease
    • /
    • v.18 no.4
    • /
    • pp.391-395
    • /
    • 2012
  • To investigate the occurrence of viruses in peach, leaf samples were collected from peach trees in commercial orchard of six areas in Korea. Reverse transcription polymerase chain reaction (RT-PCR) was used to identify the presence of the following stone fruit viruses: Apple chlorotic leaf spot virus (ACLSV), Apple mosaic virus (ApMV), Prune dwarf virus (PDV), Prunus necrotic ringspot virus (PNRSV) and Plum pox virus (PPV). About 65.0% of the 515 samples were infected with ACLSV and PNRSV. Virus-like symptoms showing mosaic on leaves was observed in ACLSV infected peach trees. However, PNRSV infected peach trees showed no symptoms. These viral DNAs by sequence analysis were confirmed 4 ACLSV isolates and 3 PNRSV isolates. The Korean peach isolates of ACLSV and PNRSV showed 70-99% and 88-99% amino acid sequence identities, respectively, with those reported previously and their amino acid sequence identities with each other were approximately 95% and 88%, respectively. Phylogenetic analysis indicated that the Korean ACLSV isolates belong to the A group of ACLSV. The Korean PNRSV isolates reported in this study were grouped into I (PV32), II (PV96) and III (PE5) groups.

Chinese yam necrotic mosaic virus Isolated from Chinese Yam in Korea (한국산 장마(Dioscorea oppasita cv. Jang-Ma)에서 분리한 Chinese yam necrotic mosaic virus)

  • Kang, Dong-Kyoon;Kondo, Toru;Shin, Jong-Hee;Shin, Hye-Young;Sung, Jung-Hyun;Kang, Sang-Gu;Chang, Moo-Ung
    • Research in Plant Disease
    • /
    • v.9 no.3
    • /
    • pp.107-115
    • /
    • 2003
  • Chinese yam (Dioscorea opposita cv. Jang-Ma) plants showing necrotic mosaic symptom were collected from their growing fields in Andong, Euisong, Gunwi and Daegu, Korea. Direct negative stainning method by electron microscope showed filamentous particles of about 660 nm in length. Immunosorbent electron microscopy (ISEM) revealed filamentous particles of 660nm decorated with antiserum of Chinese yam necrotic mosaic virus (ChYNMV). The virues purified partially were used to isolate viral RNA as template for RT-PCR to amplify about 1.2 kbp of 3'-terminal region (coat protein, 3'-UTR) with ChYNMV specific and oligo-dT primers. Amino acids sequences of amplified CP genes revealed that the viruses shared 97.9% similarity with ChYNMV (AB044386) wh ich was known as the member of Macluravirus. So the viruses from Chinese yam (D. opposita cv. Jang-Ma) plants were identified as ChYNMV. Comparing the CP amion acid sequences of ChYNMV strains with other macluraviruses such as Cardamon mosaic virus (CdMV), Narcissus latent virus (NLV) and Maclura mosaic virus (MacMV) revealed that N-terminal was the most varialbe region and conserved regions were present within the genus Macluravirus.

Abortion Caused by Bovine Brucellosis in the Chonnam Province (전남 지방의 소에서 브루셀라병에 의한 유산)

  • Oh, Hyun-Yi;Park, Chul-Ho;Kim, Sang-Il;Bae, Jae-Han;Ryu, Jae-Sun;Kim, Bang-Sil;Lee, Ju-Hwan;Kim, Jae-Pung;Park, In-Chul;Kim, Jong-Taek;Suh, Guk-Hyun;Son, Chang-Ho;Oh, Ki-Seok
    • Journal of Embryo Transfer
    • /
    • v.25 no.1
    • /
    • pp.43-50
    • /
    • 2010
  • The specimens from 32 aborted fetus and 274 aborted cows were collected in 168 farms of Chonnam province from 2005 to 2008 and were tested the brucellosis. The results obtained are summarized as follows. In the 32 aborted fetus, bovine brucellosis was detected in 12 heads (37.5%), bovine viral diarrhea-mucosal disease was detected in 7 heads (19%), ainovirus infection was detected in 1 head (3.1%), and multi-infection of BVD and brucellosis was not detected, respectively. In the 306 cases of aborted fetus and cows, bovine brucellosis was detected in 44 heads (14%). Status of abortion were confirmed in 63 farms (38%) out of 168 farms from June to August. From the point of raising scale, studies found that 128 farms (76%) out of all raised under 20 heads. The incidence of abortion by brucellosis was mainly showed in 30 heads (68.1%) about 151~250 days of gestation. In the result of the 18 farms survey, the causes of infections were detected movement of infected cattle in 5 farms (28%), unknown cause in 12 farms (67%), and recurrence in 1 farm (5%). The results of this study suggest to take an advantage of the prevention and fundamental research for bovine brucellosis in Chonnam province.

Study on Alteration of Interleukin-$1{\beta}$, -2, -6 Production and Serum Level in Schizophrenic Patients (정신분열증 환자에서 Interleukin-$1{\beta}$, -2, -6 생산능과 혈청농도 변화에 관한 연구)

  • Kim, Yong-Ku;Lee, Min-Soo;Suh, Kwang-Yoon
    • Korean Journal of Biological Psychiatry
    • /
    • v.1 no.1
    • /
    • pp.98-108
    • /
    • 1994
  • The etiology and pathophysiology of schizophrenia remain unknown. It has been postulated that infectious-autoimmune process may play a role in the pathogenesis of symptoms in some schizophrenic patients. Findings of altered interleukin(IL) regulation have been regarded as additional proof that schzophrenia has an infectious-autoimmune background. In the present study, we measured mitogen-stimulated production of and serum level of IL-$1{\beta}$, IL-2, IL-6 using ELISA in 16 neuroleptic-free schizophrenic patients and in 16 age, sex matched healthy controls. The results were as follows : 1) There was a significant decrease of IL-2 production in schizophrenic patients than in normal controls(respectively $1.90{\pm}0.13ng/m{\ell}$, $2.79{\pm}0.14ng/m{\ell}$, p<0.001). But there was no significant difference of IL-$1{\beta}$ production and IL-6 production between schizophrenic patients and normal controls. 2) There was a significant increase of serum level of IL-2 in schizophrenic pateitns than in normal controls(respectively $184.8{\pm}12.8pg/m{\ell}$, $104.2{\pm}34.2pg/m{\ell}$, p<0.01). Serum level of IL-$1{\beta}$ was partially detected in both groups and serum level of IL-6 was not detected in both groups. 3) There was no significant differences of IL-$1{\beta}$, -2, -6 production & serum level of IL-2 according to male vs female, paranoid type vs undifferentiated type, drug-naive group vs drug-free group in schizophrenic patients. 4) There was significant correlation between IL-$1{\beta}$ and IL-6 production(r=0.86, p<0.001). No correlation between IL-$1{\beta}$, -2, -6 production, serum level of IL-2 and age, duration of illness, and BPRS score was found. It has been suggested that the low lymphocyte production of IL-2 in the patients with autoimmune disease occurs because the T cells are activated and lymphocyte-derived IL-2 has been released into the serum. The authors suggest that decreased IL-2 production in our schizophrenic patients is due to increased IL-2 serum level in those patients. Thus our finding of low IL-2 production and high serum level of IL-2 in our schizophrenic patients is compatible with the possibility that our patients have an autoimmune process. Further study on relationship between IL alteration and other immunological abnormalities(the presence of serum autoantibody and of anti-brain antibody, $CD4^+$, $CD8^+$ cell index, etc) in schizophrenic patients will be warranted.

  • PDF

Mathematical Modeling of the Novel Influenza A (H1N1) Virus and Evaluation of the Epidemic Response Strategies in the Republic of Korea (수학적 모델을 이용한 신종인플루엔자 환자 예측 및 대응 전략 평가)

  • Suh, Min-A;Lee, Jee-Hyun;Chi, Hye-Jin;Kim, Young-Keun;Kang, Dae-Yong;Hur, Nam-Wook;Ha, Kyung-Hwa;Lee, Dong-Han;Kim, Chang-Soo
    • Journal of Preventive Medicine and Public Health
    • /
    • v.43 no.2
    • /
    • pp.109-116
    • /
    • 2010
  • Objectives: The pandemic of novel influenza A (H1N1) virus has required decision-makers to act in the face of the substantial uncertainties. In this study, we evaluated the potential impact of the pandemic response strategies in the Republic of Korea using a mathematical model. Methods: We developed a deterministic model of a pandemic (H1N1) 2009 in a structured population using the demographic data from the Korean population and the epidemiological feature of the pandemic (H1N1) 2009. To estimate the parameter values for the deterministic model, we used the available data from the previous studies on pandemic influenza. The pandemic response strategies of the Republic of Korea for novel influenza A (H1N1) virus such as school closure, mass vaccination (70% of population in 30 days), and a policy for anti-viral drug (treatment or prophylaxis) were applied to the deterministic model. Results: The effect of two-week school closure on the attack rate was low regardless of the timing of the intervention. The earlier vaccination showed the effect of greater delays in reaching the peak of outbreaks. When it was no vaccination, vaccination at initiation of outbreak, vaccination 90 days after the initiation of outbreak and vaccination at the epidemic peak point, the total number of clinical cases for 400 days were 20.8 million, 4.4 million, 4.7 million and 12.6 million, respectively. The pandemic response strategies of the Republic of Korea delayed the peak of outbreaks (about 40 days) and decreased the number of cumulative clinical cases (8 million). Conclusions: Rapid vaccination was the most important factor to control the spread of pandemic influenza, and the response strategies of the Republic of Korea were shown to delay the spread of pandemic influenza in this deterministic model.

Occurrence of Apple Scar Skin viroid and Relative Quantity Analysis Using Real-time RT-PCR (Apple Scar Skin viroid 발생상황 및 Real-time RT-PCR을 이용한 상대정량 분석)

  • Kim, Dae-Hyun;Kim, Hyun-Ran;Heo, Seong;Kim, Se-Hee;Kim, Min-A;Shin, Il-Sheob;Kim, Jeong-Hee;Cho, Kang-Hee;Hwang, Jeong-Hwan
    • Research in Plant Disease
    • /
    • v.16 no.3
    • /
    • pp.247-253
    • /
    • 2010
  • Apple scar skin viroid (ASSVd) is one of the smallest viral pathogens infecting fruits, especially apple, and causes a significant damage to fruit trees. ASSVd usually induced the skin-dapple ring symptoms, but in 'Fuji' varieties, corked spot were occurred on the fruit skin in 2009. This new symptom will be of great helpful to diagnosis ASSVd in sight. ASSVd was surveyed in apple and pear from 2009 to 2010 in Korea, and ASSVd was identified in 20 out of 1,193 trees. The infection rate was 1.7%. To screen the infectivity of ASSVd among apple cultivars, real-time RT-PCR was applied followed by designing of ASSVd specific primers based on highly conserved regions of several ASSVd isolates including Korean isolate. NADH dehydrogenase subunit 5 (nad 5) gene, which is mRNA of the mitochondrial gene, was used for internal control. In this study, ASSVd infected apples were classified into 12 groups depending on different symptoms and symptom severity (scaring, rusting or malformation). Taken together, this study suggested that real-time PCR analysis was more sensitive to detect the low copy of ASSVd on early viroid infected apple skins than regular RT-PCR method.

A Survey of Viral Diseases of Proso Millet (Panicum miliaceum L.) and Sorghum (Sorghum bicolor L.) in South Korea (국내 기장 및 수수의 바이러스병 발생 조사)

  • Min, Hyun-Geun;Park, Chung Youl;Lee, Hong-Kyu;Yeom, Yoon-Ah;Oh, Jonghee;Kim, Bong-Sub;Lim, Seungmo;Yoon, Youngnam;Lee, Su-Heon
    • Research in Plant Disease
    • /
    • v.23 no.3
    • /
    • pp.262-267
    • /
    • 2017
  • Throughout year 2015 to 2016, 101 proso millet and 200 sorghum samples were collected from five provinces in South Korea. The samples were subjected to paired-end RNA sequencing and further analyzed by RT-PCR. The results indicated that Rice black-streaked dwarf virus (RBSDV) was detected from sorghum collected in Gyeongsang province. The other four viruses, including RBSDV, Rice stripe virus (RSV), Barley virus G (BVG), and Cereal yellow dwarf virus (CYDV), were detected from proso millet. Among four viruses, both RSV and RBSDV were identified high frequency from proso millet collected from Gyeongsang province. Otherwise, BVG was nearly equally identified from five provinces, suggesting that the virus was supposedly widespread nationwide. RBSDV was first identified from both proso millet and sorghum in South Korea. The other virus annotated CYDV identified proso millet was shown to have relatively low identities compared to CYDV previously reported, suggesting that the virus might be new member of Polerovirus.

Construction of FMDV VP1 Gene Using Artificial DNA Synthesis and Transformation of Nicotiana tabacum Using Agrobacterium Vector System (유전자 인공합성을 이용한 구제역 유전자 VP1의 제작과 Agrobacterium Vector System을 이용한 담배 형질전환)

  • Lee, Eun-Jung;Lim, Hee-Young;Kim, Sung-Hoon;Kang, Kyung-Sun;Park, Young-Doo;Yun, Choong-Hyo;Yoon, Byoung-Su
    • Journal of Plant Biotechnology
    • /
    • v.31 no.4
    • /
    • pp.285-293
    • /
    • 2004
  • FMDV is a viral pathogen that caused foot-and-mouth disease in animals. VP1 is a major capsid protein of FMDV. It is known as one of best materials for the FMDV diagnosis and for the development of protein vaccine. In this study, 633 bp of VP1 gene was modified for the expression of VP1 in plant, based on the VP1 DNA sequence from FMDV taiwan O type and from FMDV isolated vietnam. The. deduced DNA fragment was artificially synthesized using the multiple fragment extension with long-nucleotides. A new plant transgenic vector system, pCAMBIA139011 was constructed on the basis of pBI12l and pCAMBIA1390. Using this vector system and GFP gene or modified VP1 gene, each target gene was introduced into Nicotiana tabacum. The insertion of whole target gene was successfully confirmed in each transgenic plant named GFP-A7 and VP1-4, respectively. The expression level of each gene was estimated by RT-PCR and Real-Time PCR using VP1, GFP specific primers.

A Case of Feline Proliferative Eosinophilic Keratitis Treated by Topical Cyclosporine and Corticosteroids (고양이 증식성 호산구성 각막염에서 국소적인 사이클로스포린과 코르티코스테로이드 점안 1례)

  • Ahn, Jeong-Taek;Jeong, Man-Bok;Kim, Se-Eun;Park, Young-Woo;Kim, Tae-Hyun;Ahn, Jae-Sang;Lee, So-Ra;Lee, Chang-Woo;Seo, Kang-Moon
    • Journal of Veterinary Clinics
    • /
    • v.27 no.6
    • /
    • pp.751-754
    • /
    • 2010
  • A 6-year-old spayed female Persian cat presented with a 3-month history of recurrent ulcerative keratitis with noticeable opacification and vascularization of the right cornea. The lesion was nonresponsive to topical antibiotics and to nonsteroidal anti-inflammatory drugs. Ophthalmic examination showed signs of ocular discomfort, such as epiphora and blepharospasm, in the right eye. Biomicroscopic examination revealed an irregular, edematous, vascularized mass with pink to white tissue on the entire cornea and mild conjunctivitis. A tentative diagnosis of feline proliferative eosinophilic keratitis (FPEK) was made on the basis of clinical appearance. Cytologic examination of the cornea showed a mixture of numerous eosinophils and mast cells, which confirmed the original diagnosis of FPEK. The cat was treated with a topical antibiotic-corticosteroid combination, cyclosporine ointment, trifluridine eye drops, and oral Llysine. The clinical signs improved remarkably 18 days after the cat was first examined. The short-term use of corticosteroids and long-term use of cyclosporine and an anti-viral agent resolved the lesion without recurrence of the disease for 1 year.