• Title, Summary, Keyword: spike glycoprotein gene

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Analysis of the spike glycoprotein gene and nonstructural protein gene of transmissible gastroenteritis virus using PCR and RFLP analysis (PCR과 RFLP분석을 이용한 transmissible gastroenteritis virus의 spike glycoprotein gene과 nonstructural protein gene의 분석)

  • Kwon, Hyuk-moo
    • Korean Journal of Veterinary Research
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    • v.36 no.3
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    • pp.627-633
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    • 1996
  • To analyze the genomic diversity of transmissible gastroenteritis virus (TGEV), the N-terminal half of the spike (S) glycoprotein gene and nonstructural protein gene (open reading frames 3 and 3-1) were amplified by reverse transcriptase reaction and polymerase chain reaction (RT-PCR), and analyzed using restriction fragment length polymorphism (RFLP) patterns of the amplified DNA. In this study, TGEV Miller (M6) and Purdue (P115) strains were used as reference strains, and two vaccine strains (MSV and STC3) and four Korea isolates (P44, VRI-WP, VRI-41, and VRI-48) were analyzed. All TGEV strains were amplified with three TGEV primer pairs. Although there was some exception in RFLP analysis, this method differentiated TGEV strains into following groups : Miller group (M6 and MSV), Purdue group (PUS, STC3, P44, VRI-WP, VRI-41, and VRI-48). Using Sau3AI and SspI, VRI-48 was differentiated from the Miller and Purdue type viruses. The RT/PCR in conjuction with RFLP analysis was a rapid and valuable tool for differentiating several strains of TGEV. This study revealed the occurences of distinct difference in genome of TGEV strains.

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Sequence of the spike gene containing antigenic sites A and D of transmissible gastroenteritis virus isolated in Korea (국내분리 돼지 전염성 위장염 바이러스의 antigenic site A와 D를 포함하는 spike gene의 염기서열 분석)

  • Kwon, Hyuk-moo;Pi, Jae-ho;Seong, Hwan-woo
    • Korean Journal of Veterinary Research
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    • v.38 no.2
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    • pp.319-327
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    • 1998
  • The nucleotide sequences of spike (S) glycoprotein containing antigenic sites A and D of TGEV isolated in Korea were determined and compared with published sequences for TGEVs. The TGEV 133 and DAE5 strains had 97.40% nucleotide sequence similarity. The overall nucleotide sequence similarity of the 133 and DAE5 strains compared with other TGEV strains was between 96.86% and 99.15%. The similarity of the predicted amino acid sequence of the 133 and DAE5 strains was 94.93%. The TGEV 133 and DAE5 strains had 94.93-98.61% amino acid similarity with published sequences of other TGEV strains. The sequences of amino acid codons in the antigenic sites A and D were identical among all the viruses although there were several nucleotide changes in region containing antigenic sites A and D of Korean TGEV isolates. By phylogenetic analysis of the sequences, two Korean isolates 133 and DAE5 seemed to be derived from different lineages. These studies showed that a distinct difference in genome exists among TGEV field strains isolated in Korea.

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Genetic sequence and phylogenetic analysis of spike genes of Porcine epidemic diarrhea virus (PEDV) in Chung-Nam area (충남지역 돼지유행성설사 바이러스 Spike 유전자 염기서열 및 계통분석)

  • Park, Hyo-Seon;Yook, Sim-Yong;Jeon, Dong-Min;Lee, Jin-Ju;Shin, Chang-Ho
    • Korean Journal of Veterinary Service
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    • v.39 no.4
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    • pp.259-266
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    • 2016
  • Porcine epidemic diarrhea virus (PEDV) causes an acute and lethal watery diarrhea in piglets that is great economic losses to the swine country worldwide. The spike (S) glycoprotein is an important determinant for PEDV biological properties. In the present study, we determined the full-length S gene sequences of five Chung-nam PEDV field isolates collected in 2016. The S gene was amplified by RT-PCR, purificated, sequenced, analyzed and then compared with published sequences of other PEDV strains. 5 field strains share 98.5%~99.9% homologies with each other at the nucleotide sequence level and 96.7%~99.9% homologies with each other at the amino acids sequence level. Most field strains have nucleotide insertions, deletions and mutation regions, and show lower homologies (93.1~93.8%) with classical and vaccine strains, however higher homologies (99.1%~99.5%) with US PEDV isolates in 2013. By phylogenetic tree analysis based on nucleotide sequence, five PEDV field isolates were clustered into Genogroup 2b but differ genetically from the vaccine strains (SM-98 and DR-13).

Genetic characterization and phylogenetic analysis of infectious bronchitis virus isolated in Jeonbuk (전북지역 전염성기관지염 바이러스의 유전적 특성)

  • Chu, Keum-Suk;Kim, Ji-Hyun;Lee, Jeong-Won;Choi, Kwang-Lim
    • Korean Journal of Veterinary Service
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    • v.38 no.1
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    • pp.51-56
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    • 2015
  • Infectious bronchitis virus (IBV) causes an acute and highly contagious viral disease of chicken that is great economic losses to the poultry industry worldwide. Among the IBV structural proteins, the high rate spike glycoprotein S1 gene mutation and antigenic variant strains have been reported in many countries. During the years 2012~2014, 10 IBV strains were isolated from infected chicken farms distributed in provinces of Jeonbuk. Analysis of the S1 gene sequences amplified from 10 isolated strains with QX strains showed nucleotide homologies ranging from 96.5 to 95.4%. Phylogenetic analysis revealed that all strains were clustered into QX-like groups. This study suggests that QX-like IBVs are circulating in commercial chicken farms in Jeonbuk. Therefore, the continuing survellance is significantly important for prevention and control of BIV infection.

Molecular biological characterization of transmissible gastroenteritis viruses isolated in Korea (돼지 전염성 위장염 바이러스(국내분리주)의 분자생물학적 특성 규명)

  • Kwon, Hyuk-moo;Pi, Jae-ho
    • Korean Journal of Veterinary Research
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    • v.38 no.2
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    • pp.304-313
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    • 1998
  • Sixteen Korean field transmissible gastroenteritis viruses (TGEVs) were isolated using swine testicular cell (STC) and the genomic diversity of them was analyzed. All TGEV isolates produced a typical cytopathic effect in STC and were confirmed as TGEV by immunofluorescence assay using monoclonal antibody against TGEV and PCR using TGEV specific primers. RNAs from TGEV field isolates and vaccine TGEV were extracted and amplified by RT and PCR. The RT-PCR products were digested with selected restriction enzymes and analyzed RFLP patterns. The N-terminal end region of S gene and ORF 3 and 3-1 genes of TGEV amplified by TGEV specific primer pairs seemed to be conserved. Most specific variations were detected in S gene amplified by TGEV 4/6 primer pairs which includes antigenic sites A and D. When the PCR products were treated with Sau3AI and Ssp I, Bvac(vaccine strain), field isolates 133 and 347 were differentiated from Miller and Purdue types. In the case of D5 field isolates, it was classified into Purdue type by Sau 3AI but classified into independent TGEV by Ssp I. Two different TGEV strains from D2 sample were confirmed by plaque purification and RT-PCR-RFLP analysis. To investigate the change occurring in TGEV genome after serial passage, the TGEV P44 strain was passaged through STC. There were specific changes in S gene and a large deletion was observed in ORF 3 and 3-1 genes. These studies showed that a distinct difference in genome exists among TGEV field isolates.

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Pathogenesis of infectious bronchitis virus with different routes of inoculation and the effect of in vivo serial passage in nephropathogenicity using cloacal infection

  • Lee, Chang-Won
    • Korean Journal of Veterinary Service
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    • v.25 no.1
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    • pp.87-96
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    • 2002
  • In this study, we wanted to determine if the respirotropic JMK strain of infectious bronchitis virus(IBV), which has a spike glycoprotein gene that is 99% similar to the nephropathogenic Gray strain of IBV, could adapt and cause lesions in the kidney following intracloacal passage in chickens. Two day old specific pathogen free(SPF) cchickens were infected with Gray and JMK strains by the intraocular and cloacal route. Several tissue samples were collected at various times. Viruses were recovered from more tissues and earlier in the infection from chickens infected cloacally than chickens infected intraocularly. Virus was isolated from the kidney of chickens infected with Gray by the intraocular route and JMK by the intracloacal route, but not from chicken given JMK the intraocular route. Histopathologically, interstitial nephritis was observed in Gray infected chickens. However, viral RNA or antigen were not detected in the kidney by in situ hybridization and immunohistochemistry. We further passaged the JMK strain ten times in two day old SPF chickens using cloacal inoculation. We examined the virus titer and histopathological change in the kidney at each passage level. The amount of virus recovered from the kidney was stable throughout this serial passage and the passaged virus did not caused renal damage. Further, virus could not be isolated from the kidney when chickens were infected with the passaged virus by the intraocular route. We conclude that the JMK strain has a strict upper respiratory tract tropism since cloacal passage did not produce nephrotropism or nephropathogenicity.