• Title/Summary/Keyword: VP8

Search Result 271, Processing Time 0.032 seconds

Use of the Synthetic Gene Encoding the Truncated Human Rotavirus VP8* Protein in Escherichia coli for Production of Vaccine Candidates or Development of Diagnostic Antibodies (합성 유전자를 이용하여 Escherichia coli에서 백신 후보의 생산 혹은 진단용 항체의 개발을 위한 인간 rotavirus VP8* 부분 단백질의 발현)

  • Kim, Sang-Rae;Lee, Bheong-Uk
    • Journal of Life Science
    • /
    • v.28 no.4
    • /
    • pp.478-482
    • /
    • 2018
  • Human rotavirus is a causative agent of acute diarrhea among children. The artificial gene encoding the truncated $VP8^*$ protein of human rotavirus A (serotype 1 strain WA) was synthesized according to the Escherichia coli codon preference. The synthetic $VP8^*$ gene also possessed the NdeI and HindIII restriction sites for the convenient in-frame cloning for translation and a 6-histidine tag at C-terminus for Ni+ affinity purification. Molecular weight of the truncated $VP8^*$ protein deduced from the nucleotide sequences of the artificial gene was a 19.7-kDa. This synthetic $VP8^*$ DNA fragment was inserted into the pT7-7 expression vector and transformed into E. coli BL21 (DE3). Transformants harboring the synthetic gene encoding the $VP8^*$ protein was induced by supplement of a final concentration of 0.05 mM ITPG at $20^{\circ}C$. Protein crude extract from the E. coli transformants was subjected to Western blotting with the mouse anti-rotavirus capsid antibody, showing ~20-kDa $VP8^*$ protein band. The truncated $VP8^*$ protein band was also observed by Western blotting using the rabbit polyclonal antibody serum made against the truncated $VP8^*$ protein. This study suggested that the synthetic gene could be used as an easy way to produce the antigenic vaccine candidate for control of virus-associated diseases or to develop antibodies for diagnostic purpose.

Optimal Exposure Conditions according to Detector Type in Chest Digital Radiography (디지털흉부X선촬영에서 검출기 방식에 따른 최적의 노출조건)

  • Lee, Won-Jeong
    • Journal of the Korea Safety Management & Science
    • /
    • v.17 no.4
    • /
    • pp.213-221
    • /
    • 2015
  • The aim of this study was to set up the optimal exposure condition according to detector type considering image quality (IQ) with radiation dose in chest digital radiography. We used three detector type such as flat-panel detector (FP) and computed radiography (CR), and charge-coupled device (CCD). Entrance surface dose (ESD) was measured at each exposure condition combined tube voltage with tube current using dosimeter, after attaching on human phantom, it was repeated 3 times. Phantom images were evaluated independently by three chest radiologists after blinding image informations. Standard exposure condition using each institution was 117 kVp-AEC at FP and 117 kVp-8 mAs at CR, and 117 kVp-8 mAs at CCD. Statistical analysis was performed by One way ANOVA (Dunnett T3 test) using SPSS ver. 19.0. In FP, IQ scores were not significant difference between 102 kVp-4 mAs and 117 kVp-AEC (28.4 vs. 31.1, p=1.000), even though ESD was decreased up to 50% ($62.3{\mu}Gy$ vs. $125.1{\mu}Gy$). In CR, ESD was greatly decreased from 117 kVp-8 mAs to 90 kVp-8 mAs without significant difference of IQ score (p=1.000, 24.6 vs. 19.5). In CCD, IQ score of 117 kVp-8 mAs was similar with 109 kVp-8 mAs (29.6 vs. 29.0), with decreasing from $320.8{\mu}Gy$ to $284.7{\mu}Gy$ (about 11%). We conclude that optimal x-ray exposure condition for chest digital radiography is 102 kVp-4 mAs in FP and 90 kVp-8 mAs in CR, and 109 kVp-8 mAs in CCD.

Production and diagnostic application of monoclonal antibodies against infectious bursal disease virus (IBDV에 대한 단크론항체 생산 및 진단적 응용)

  • Ryu, Min-Sang;Song, Yoon-Ki;Lee, Seung-Chul;Mo, In-Pil;Kang, Shien-Young
    • Korean Journal of Veterinary Service
    • /
    • v.34 no.1
    • /
    • pp.5-12
    • /
    • 2011
  • Infectious bursal disease (IBD) caused by infectious bursal disease virus (IBDV) is a highly contagious viral disease in chicken. It causes heavy economic loss by immune suppression and high mortality. The IBDV, designated Avibirnavirus in the Family Birnaviridae, has a double-stranded RNA genome formed by two segments, segment A and segment B. Segment A encodes a 108 KDa polypeptide that is self-cleaved to produce pVP2, VP3 and VP4, and later pVP2 is cleaved to VP2. The VP2 contains the antigenic regions responsible for elicitation of neutralizing antibodies and VP3 is a major immunogenic protein of IBDV. In this study, monoclonal antibodies (MAbs) specific for IBDV were produced and characterized. All 15 MAbs were specific for IBDV and did not react with other viruses used in this study. The protein specificity of MAbs was determined by comparing the reactivity patterns of each MAb with IBDV VP2 and VP234 recombinant baculoviruses and Western blot analysis. As a result, 7 MAbs (1F5, 2C8, 2F4, 3C7, 4C3, 6F11, 6G5) and 5 MAbs (2A4, 2G2, 3F5, 3G2, 4F10) were specific for VP2 and VP3, respectively. The protein specificity of 3 MAbs (2B8, 3F7, 3F8) were not determined. Five (2C8, 2F4, 4C3, 6F11, 6G5) of the VP2-specific MAbs had a neutralizing activity against IBDV. Some MAbs reacted with IBDV-infected bursa of Fabricius by indirect fluorescence antibody (IFA) and immunohistochemistry (IHC) assay. The MAbs produced in this study would be used for diagnostic reagents for the detection of IBDV infection.

Vaccination of Shrimp (Litopenaeus vannamei) against White Spot Syndrome Virus (WSSV) by Oral Vaccination of Recombinant Fusion Protein, rVP19+28 (사료급이(oral feeding)에 의한 vaccination을 통한 흰반점바이러스(WSSV)에 대한 재조합단백질 rVP19+28의 백신효능의 확인)

  • Nguyen, Thi-Hoai;Kim, Yeong-Jin;Choi, Mi-Ran;Kim, Sung-Koo
    • Journal of Life Science
    • /
    • v.20 no.8
    • /
    • pp.1181-1185
    • /
    • 2010
  • This study was carried out to evaluate the vaccination effects of recombinant fusion protein rVP19+28 against WSSV in shrimp, Litopenaeus vannamei. The VP19+28 gene fused with VP19 and VP28 genes was inserted into pET-28a(+) expression vector and cloned in E. coli BL21 (DE3) to produce fused gene product recombinant VP19+VP28 as a single protein. For the vaccination, the shrimps were fed with pellets coated with purified recombinant protein, rVP19+28, for 2 weeks. Then, constant amounts of WSSV at $1{\times}10^2$ diluted stocks were injected to the muscle of the shrimp for the in vivo challenge tests. Non-vaccinated shrimps showed a cumulative mortality of 100% at 11 days post-challenge. The shrimps vaccinated with the inactivated E. coli BL21 as a host cell control showed cumulative mortality of 100% at 17 days post-challenge. The shrimps vaccinated with rVP19, rVP28 and rVP19+28 showed mortalities of 66.7%, 41.7% and 41.7% at 21 days post-challenge, respectively. These results indicated that the rVP28 and rVP19+28 had relatively high vaccination effects against WSSV infection. However, this study suggests that the fusion protein rVP19+28 was more effective for the protection of shrimp against WSSV than rVP28, even though the cumulative mortalities were the same 21 days post-challenge.

Quality and Shelf-life of Chilled Smoked Pork Belly Depending on Packaging Methods (포장방법에 따른 냉장 훈제삼겹살의 품질과 저장성)

  • Lim, Ji-Hoon;Cheong, Sung-Hee;Lee, Sung-Ki;Lee, Keun-Taik
    • KOREAN JOURNAL OF PACKAGING SCIENCE & TECHNOLOGY
    • /
    • v.16 no.2_3
    • /
    • pp.43-51
    • /
    • 2010
  • The changes in quality and shelf life of smoked pork belly according to packaging methods were investigated during storage at $5{\pm}1^{\circ}C$ up to 12 days. Three packaging treatments including air-containing packaging (AC), vacuum packaging (VP), and packaging incorporated with oxygen scavenger (OS) were applied in this experiment. In all treatments, the initial total aerobic plate count (TPC) was 2.4 log cfu/g which was increased with storage period. The rapidest increase of TPC was observed in the AC samples, followed by OS and VP. The VP samples showed inhibiting effects on the growth of Pseudomonas spp. and coliforms over the storage. The TBA values were increased in the order of AC-OS-VP. The VBN values of the VP and OS samples tended to be increased slower than the AC samples during the storage. According to the sensory evaluations, the point of losing marketability in the raw AC, OS, and VP samples was determined to be the day 8, 10, and 12, respectively. However, the evaluations for the cooked samples showed the AC and OS samples preserved the marketing value until the day 10 and 12, while the VP samples until the day 12 except in the parameter of texture. In conclusion, it was found that the most appropriate packaging method for preserving the quality and extending the shelf life of chilled smoked pork belly is vacuum packaging.

  • PDF

Evaluation on Usefulness of BMI Application to Urological CT Examination (비뇨기계 CT 검사 시 체질량 지수 적용의 유용성 평가)

  • Kim, Hyeon-jin;Im, In-chul
    • Journal of the Korean Society of Radiology
    • /
    • v.12 no.2
    • /
    • pp.185-191
    • /
    • 2018
  • The purpose of this study was to evaluate the usefulness of BMI application to Urography CT by applying different tube voltages in accordance with body mass index. Group A (n = 38) with body mass index of lower than 25 was examined with tube voltage of 100 kVp while Group B (n = 45) with a BMI of 25 and higher was examined with tube voltage of 120 kVp. C group (n = 37) with body mass index (BMI) of lower than 25 was examined with tube voltage of 120kVp. Although the difference in average dose between group A (100 kVp) and group C (120 kVp) with low body mass index (BMI) of lower than 25 was $214.8mGy{\cdot}cm$, there was no significant difference in qualitative evaluation and, compared with patient group with body mass index of 25 and higher, results obtained were rather good. Therefore, this study verified that the tube voltage of lower than 100 kVp does not have adverse effect on the quality of image for patients with body mass index (BMI) of lower than 25.

Response of Electrocardiogram of Nile tilapia, Oreochromis niloticus to Electric Stimulus (전기자재에 대한 역돔의 심전도)

  • 한규환;양용림
    • Journal of the Korean Society of Fisheries and Ocean Technology
    • /
    • v.38 no.4
    • /
    • pp.278-283
    • /
    • 2002
  • The response of electrocardiogram(ECG) of Nile tilapia, Oreochromis niloticus [Linnaeus] was studied to the electric stimulus which was given to a certain part of body The experiments were performed in such a way that three levels of electric stimulus (20, 30, 40 Vp ; 10 msec) were given to fishes with electrode inserted into their bodies and then their ECGs were recorded continuously for 60 minutes in the water temperature of 16~18$^{\circ}C$ The results of the experiments were divided by day and night, and then were analyzed by experimental conditions as follows; 1. Nile tilapia reached a stable condition within 3 minutes after the electrode inserted into their bodies during anesthesia. In stable condition, the heart rates average was 45.8 beat/min during daytime and 45.0 beat/min at night. The action potentials average was 1.76 $mutextrm{V}$during daytime and 1.75 $mutextrm{V}$ at night. 2. The heart rates average by three levels of electric stimulus were \circled1 In the stimulus condition, the heart rates were 34.9 beat/min during daytime and 33.4 beat/min at night for the 20 Vp level, 36.8 bea/min during daytime and 36.0 beat/min at night for the 30 Vp level, and 38.0 beat/min during daytime and 36.4 beat/min at night for the 40Vp level. \circled2 In the recovery condition, the action potentials were 45.5 beat/min during daytime an 45.1 beat/min at night for the 20Vp level, 47.9 beat/min during daytime and 49.0 beat/min at night for the 30Vp level, and 51.4 beat/min during daytime and 50.7 beat/min at night for the 40Vp level 3. The action potentials average by three levels of electric stimulus were, \circled1 In the stimulus condition, action potentials were 2.54 $mutextrm{V}$ during daytime and 2.39 $mutextrm{V}$ at night for the 20 Vp level, 3.30 $mutextrm{V}$ during daytime and 2.30 $mutextrm{V}$ at night for the 30 Vp level and 6.05 $mutextrm{V}$ during daytime and 3.23 $mutextrm{V}$ at night for the 40 Vp level. \circled2 In the recovery condition, action potentials were 1.92 $mutextrm{V}$ during daytime and 1.95 $mutextrm{V}$ at night for the 20 Vp level and 2.78 $mutextrm{V}$ during daytime and 2.21 $mutextrm{V}$ at night for the 30Vp level and 3.6 0 $mutextrm{V}$ during daytime and 2.98 $mutextrm{V}$ at night for the 40 Vp level.

Distribution of Disease Severity and Group A Rotavirus Genotypes (VP7 & VP4) in Children with Acute Rotavirus Gastroenteritis (급성 로타바이러스 장염 소아에서 질환의 중증도와 로타바이러스 VP7 & VP4 유전형의 분포에 대한 연구)

  • Oh, Hyun-Ju;Kang, Hyun-Sik;Kang, Ki-Soo;Kim, Youn-Woo;Hong, Jung-Yun;Shin, Kyung-Sue;Lee, Jin-Sook;Lee, Su-Hyun;Lee, Keun-Hwa;Cho, Moon-Je;Suh, Dong-In
    • Pediatric Gastroenterology, Hepatology & Nutrition
    • /
    • v.14 no.2
    • /
    • pp.148-154
    • /
    • 2011
  • Purpose: We aimed to study the distribution of rotavirus genotypes (VP7 and VP4) and disease severity of rotavirus gastroenteritis prevalent in our community. Methods: Stool samples were collected from 156 children who were hospitalized with rotavirus gastroenteritis from December 2007 to June 2008. The disease severity of all patients was scored using the Vesikari scale. After extraction of ds-RNA of the rotavirus, cDNA synthesis using reverse transcription and polymerase chain reaction (RT-PCR) and multiplex PCR was performed. Following this, the final identification of genotypes was performed. Results: Of the 156 samples, VP7(G) and VP4(P) genotypes were identified in 147 (94.2%) and 140 (89.7%) samples, respectively. G1 (116 of 147 samples; 78.9%) and P[8] (137 of 140 samples; 97.9%) were the most prevalent, respectively. Of the 138 samples identified of combination types of VP7 and VP4, G1P[8] (111 samples; 80.4%) was the most prevalent. Other combination types varied with very low distribution rates. 9.4% of genotypes were not included in the new vaccines. The disease severity score was $11.8{\pm}3.3$ ($mean{\pm}2SD$). The distribution of disease severity was mild or moderate in 37.8% and severe in 62.2% of patients. Conclusion: The most prevalent genotype combination of rotavirus was G1P[8] and genotypes not included in the vaccines represented 9.4% in our community. Disease severity distribution of hospitalized children with rotavirus gastroenteritis was higher in the severe than in the mild and moderate categories.

Molecular characterization of avian rotavirus isolated in Korea

  • Wang, Jun-Hui;Koo, Bon-Sang;Mo, In-Pil;Kang, Shien-Young
    • Korean Journal of Veterinary Service
    • /
    • v.36 no.1
    • /
    • pp.23-30
    • /
    • 2013
  • An avian rotavirus (AvRV-2) was isolated from feces of broilers suffering from acute gastroenteritis in 2011. It was the first avian rotavirus isolated in Korea. To investigate the molecular characteristics of AvRV-2, the VP4, VP6, VP7 and NSP4 gene nucleotide sequences were determined and compared with those of rotavirus strains available in the GenBank database. The phylogenetic tree of VP7 gene showed that AvRV-2 had a high degree of nucleotide sequence homology (93.4% to 94.7%) with those of rotaviruses belonging to genotype G19 cluster. The phylogenetic tree of the VP4 gene revealed a high degree of nucleotide sequence homology (95.8% to 95.9%) with genotype P[30] rotaviruses isolated from chickens. The VP6 and NSP4 gene nucleotide sequences showed the highest identities with those of avian strains with 95.3% to 96.4% and 90.3% to 92.2%, respectively. Genetic characterization of the VP4, VP6, VP7 and NSP4 showed that AvRV-2 strain was most closely related to chicken rotavirus strains from Germany and Japan. Comparative nucleotide sequences and phylogenetic analysis indicated that avian rotavirus isolated from broilers belonged to genotype G19P[30] and it was the first report on avian rotavirus infection in Korea.

Studies on the VP4 and VP7 Genes of Bovine Rotaviruses from Field Samples Using RT-PCR and RFLP Analysis (RT-PCR과 RFLP법을 이용한 국내 소 로타바이러스 VP4 및 VP7 유전자의 특성 규명)

  • Jeon, Seong-Jin;Chang, Chung-Ho;Chung, Chung-Won;Kim, Won-Yong;Kang, Shien-Young
    • The Journal of Korean Society of Virology
    • /
    • v.28 no.2
    • /
    • pp.165-174
    • /
    • 1998
  • Characterizations of the VP4 (P type) and VP7 (G type) genes of Korean isolates of bovine rotavirus were performed using RT-PCR/RFLP and nucleotide sequencing analysis. After RT-PCR amplification of partial length (1094bp) of the VP4 and full length (1062bp) of the VP7 genes, amplified PCR products were digested with restriction endonucleases and digestion patterns were compared with those of reference rotaviruses. With the VP4 genes, four RFLP (A-D) profiles were observed; three (A, Band C) were the same as those of bovine rotavirus NCDV (P[1]), IND (P[5]) and B223 (P[11]), respectively. Profile D was the same as that of porcine rotavirus OSU (P[7]). With the VP7 genes, five RFLP profiles (I-V) were observed; three of them (I, II and III) were the same as those of bovine rotavirus NCDV (G6), Cody 1-801 (G8), and B223 (G10), respectively. Profile IV and V were atypical to those of reference bovine rotaviruses used in this study. These two profiles were identified as G6 and G5, respectively, after analyzing and comparing the nucleotide sequences. The G typing analysis revealed that 61.9% (26/42) were G6, which included G6 subtype; 28.6% (12/42) were G5; 7.1% (3/42) were G10; 2.4% (1/42) were G8. The P typing analysis revealed that 54.8% (23/42) were P[5]; 28.6% (12/42) were P[7]; 11.8% (5/42) were [11]; 4.8% (2/42) were P[1]. Our results showed that G6/P[5] were the most prevalent rotaviruses in diarrheic calves in Korea. Also, this is the first report that G5/P[7] rotaviruses were identified from cattle with diarrhea.

  • PDF