• Title/Summary/Keyword: virus mutation

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Study on potential environmental risk of virus resistant LM plants using co-inoculation of Zucchini green mottle mosaic virus (ZGMMV) and Cucumber mosaic virus (CMV) (ZGMMV와 CMV 동시 접종을 통한 바이러스 저항성 LM 식물의 잠재적 환경 위해성 연구)

  • Song, Hae-Ryong;Kim, Taesung;Kim, Sun-Jung;Kim, Yong-Hyun;Kim, Ki-Jeong;Chung, Hyen-Mi;Choi, Hee Lak;Yoon, Junheon
    • Journal of Environmental Impact Assessment
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    • v.22 no.2
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    • pp.125-134
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    • 2013
  • Plant virus coat (CP) gene-mediated protection is one of the best known approaches to protect against virus resistant transgenic plants. Transgenic N. benthamiana plants containing the CP gene of Zucchini green mottle mosaic virus (ZGMMV) were used for the environmental risk assessment of the living modified (LM) plants with plant virus resistance. The most optimal co-infection method of both ZGMMV and CMV (Cucumber mosaic virus) on Non-LM and CP-expressing LM tobacco plants was established and co-infection of CMV and ZGMMV was confirmed by polymerase chain reaction (PCR). To address the effects of LM tobacco plants on the mutation of the virus, in-vitro transcripts of CP and Replicase (Rep) derived from CMV and/or ZGMMV were inoculated onto Non-LM or LM tobacco plants. Mutation frequency of CP and Rep from CMV and ZGMMV was examined through six serial passages in Non-LM and LM tobacco plants. Little actual frequency of mutation was estimated, probably due to the limited number of transgenic plants tested in this study. However, it does not suggest environmental safety of these CP-mediated LM plants. Further study at a larger scale is needed to evaluate the environmental risk associated with the CP-expressing LM plants.

A Case Report of HBsAg Seroclearance in Chronic Hepatitis B Patient

  • Lee, Hyeok Jae;Lee, Min-Hyeok
    • Korean Journal of Clinical Laboratory Science
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    • v.44 no.3
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    • pp.142-146
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    • 2012
  • Hepatitis B surface antigen (HBsAg) seroclearance is a rare event in chronic hepatitis B virus (HBV) infection which acquires the disease early in life. A case study have examined with asymptomatic chronic hepatitis B carrier who exhibits HBsAg seroclearance in anti-HBe positive. We comprehensively studied the biochemical, virological and clinical aspects of a patient with HBsAg seroclearance. Liver biochemistry, serological markers, serum HBV DNA levels, and development of clinical complications were monitored. Mutation of hepatitis B virus is suspected serum HBsAg detected by the HBsAg assay systems of VITROS (OrthoClinical Diagnostics, USA), AxSYM (Abbott Laboratories, USA), Elecsys (Roche Diagnostics, Germany) and ADVIA Centaur (Bayer Diagnostics, USA). These four immunoassays showed negative results. Also, the patient had undetectable serum HBV DNA. Therefore, no mutation within the "a" determinant of HBsAg, which might escape detection from HBsAg immunoassay were found. Natural seroclearance was confirmed.

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Establishment of multiplex RT-PCR for differentiation between rabies virus with and that without mutation at position 333 of glycoprotein

  • Yang, Dong-Kun;Kim, Ha-Hyun;Lee, Siu;Yoo, Jae-Young
    • Journal of Veterinary Science
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    • v.21 no.2
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    • pp.22.1-22.9
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    • 2020
  • Rabid raccoon dogs (Nyctereutes procyonoides koreensis) have been responsible for animal rabies in South Korea since the 1990s. A recombinant rabies vaccine strain, designated as ERAGS, was constructed for use as a bait vaccine. Therefore, new means of differentiating ERAGS from other rabies virus (RABV) strains will be required in biological manufacturing and diagnostic service centers. In this study, we designed two specific primer sets for differentiation between ERAGS and other RABVs based on mutation in the RABV glycoprotein gene. Polymerase chain reaction analysis of the glycoprotein gene revealed two DNA bands of 383 bp and 583 bp in the ERAGS strain but a single DNA band of 383 bp in the field strains. The detection limits of multiplex reverse transcription polymerase chain reaction (RT-PCR) were 80 and 8 FAID50/reaction for the ERAGS and Evelyn-Rokitnicki-Abelseth strains, respectively. No cross-reactions were detected in the non-RABV reference viruses, including canine distemper virus, parvovirus, canine adenovirus type 1 and 2, and parainfluenza virus. The results of multiplex RT-PCR were 100% consistent with those of the fluorescent antibody test. Therefore, one-step multiplex RT-PCR is likely useful for differentiation between RABVs with and those without mutation at position 333 of the RABV glycoprotein gene.

Development of an attenuated vaccine strain from a korean respiratory type infectious bronchitis virus (한국호흡기형 닭전염성기관지염 생독백신주의 작성)

  • Choi, Kang-Seuk;Jeon, Woo-Jin;Lee, Eun-Kyoung;Kye, Soo-Jeong;Park, Mi-Ja;Kwon, Jun-Hun
    • Korean Journal of Veterinary Research
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    • v.51 no.3
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    • pp.193-201
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    • 2011
  • An attenuated vaccine strain AVR1/08 of Korean respiratory type of infectious bronchitis virus (IBV) was developed by 89th passages of IBV D85/06 strain in chicken eggs. The AVR1/08 strain had higher virus titer at least 20 times ($10^{1.3}$) than the parent virus D85/06 by egg inoculation method. The AVR1/08 strain had a single point mutation (S to Y) at position 56 of spike protein of IBV compared to parent virus IBV D85/06 strain. The mutation was observed consistently at viruses after 47th passage in chicken eggs. The AVR1/08 strain showed no virulence even after 6 passages in chickens and all chickens inoculated induced anti-IBV antibody 14 days after vaccination. The AVR1/08 strain had broad protective efficacy against QX type Korean nephropathogenic virus (Q43/06 strain), KM91 type Korean nephropathogenic virus (KM91 strain) and Korean respiratory virus (D85/06 strain). In contrast, Massachusetts (Mass) type attenuated vaccine strain H120 showed protection of 37.5 to 50% against these three viruses. Our results indicate that the AVR1/08 strain has potential as an attenuated vaccine effective in controlling IBVs circulating in Korea.

A Comparison between Low- and High-Passage Strains of Human CytomegalovirusS

  • Wang, Wen-Dan;Lee, Gyu-Cheol;Kim, Yu Young;Lee, Chan Hee
    • Journal of Microbiology and Biotechnology
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    • v.26 no.10
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    • pp.1800-1807
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    • 2016
  • To understand how human cytomegalovirus (HCMV) might change and evolve after reactivation, it is very important to understand how the nucleotide sequence of cultured HCMV changes after in vitro passaging in cell culture, and how these changes affect the genome of HCMV and the consequent variation in amino acid sequence. Strain JHC of HCMV was propagated in vitro for more than 40 passages and its biological and genetic changes were monitored. For each passage, real-time PCR was performed in order to determine the genome copy number, and a plaque assay was employed to get virus infection titers. The infectious virus titers gradually increased with passaging in cell culture, whereas the number of virus genome copies remained relatively unchanged. A linear correlation was observed between the passage number and the log10 infectious virus titer per virus genome copy number. To understand the genetic basis underlying the increase in HCMV infectivity with increasing passage, the whole-genome DNA sequence of the high-passage strain was determined and compared with the genome sequence of the low-passage strain. Out of 100 mutations found in the high-passage strain, only two were located in an open reading frame. A G-T substitution in the RL13 gene resulted in a nonsense mutation and caused an early stop. A G-A substitution in the UL122 gene generated an S-F nonsynonymous mutation. The mutations in the RL13 and UL122 genes might be related to the increase in virus infectivity, although the role of the mutations found in noncoding regions could not be excluded.

De novo genome assembly and single nucleotide variations for Soybean yellow common mosaic virus using soybean flower bud transcriptome data

  • Jo, Yeonhwa;Choi, Hoseong;Kim, Sang-Min;Lee, Bong Choon;Cho, Won Kyong
    • Journal of Applied Biological Chemistry
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    • v.63 no.3
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    • pp.189-195
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    • 2020
  • The soybean (Glycine max L.), also known as the soya bean, is an economically important legume species. Pathogens are always major threats for soybean cultivation. Several pathogens negatively affect soybean production. The soybean is also known as a susceptible host to many viruses. Recently, we carried out systematic analyses to identify viruses infecting soybeans using soybean transcriptome data. Our screening results showed that only few soybean transcriptomes contained virus-associated sequences. In this study, we further carried out bioinformatics analyses using a soybean flower bud transcriptome for virus identification, genome assembly, and single nucleotide variations (SNVs). We assembled the genome of Soybean yellow common mosaic virus (SYCMV) isolate China and revealed two SNVs. Phylogenetic analyses using three viral proteins suggested that SYCMV isolate China is closely related to SYCMV isolates from South Korea. Furthermore, we found that replication and mutation of SYCMV is relatively low, which might be associated with flower bud tissue. The most interesting finding was that SYCMV was not detected in the cytoplasmic male sterility (CMS) line derived from the non-CMS line that was severely infected by SYCMV. In summary, in silico analyses identified SYCMV from the soybean flower bud transcriptome, and a nearly complete genome of SYCMV was successfully assembled. Our results suggest that the low level of virus replication and mutation for SYCMV might be associated with plant tissues. Moreover, we provide the first evidence that male sterility might be used to eliminate viruses in crop plants.

Hepatitis B Virus Genetic Variation and TP53 R249S Mutation in Patients with Hepatocellular Carcinoma in Thailand

  • Thongbai, Chureeporn;Sa-nguanmoo, Pattaratida;Kranokpiruk, Pavanrat;Poovorawan, Kittiyod;Poovorawan, Yong;Tangkijvanich, Pisit
    • Asian Pacific Journal of Cancer Prevention
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    • v.14 no.6
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    • pp.3555-3559
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    • 2013
  • Chronic hepatitis B virus (HBV) infection and dietary exposure to aflatoxin B1 (AFB1) are major risk factors for hepatocellular carcinoma (HCC). The aim of this study was to evaluate the role of HBV genetic variation and the R249S mutation of the p53 gene, a marker of AFB1-induced HCC, in Thai patients chronically infected with HBV. Sixty-five patients with and 89 patients without HCC were included. Viral mutations and R249S mutation were characterized by direct sequencing and restriction fragment length polymorphism (RFLP) in serum samples, respectively. The prevalences of T1753C/A/G and A1762T/G1764A mutations in the basal core promotor (BCP) region were significantly higher in the HCC group compared to the non-HCC group. R249S mutation was detected in 6.2% and 3.4% of the HCC and non-HCC groups, respectively, which was not significantly different. By multiple logistic regression analysis, the presence of A1762T/G1764A mutations was independently associated with the risk of HCC in Thai patients.

Comparison of Full Genome Sequences Between Two Hepatitis B Virus Strains With or Without preC Mutation (A1896) from a Single Korean Hepatocellular Carcinoma Patient

  • Kim, Hong;Jee, Young-Mee;Mun, Ho-Suk;Song, Byung-Cheol;Park, Joo-Hee;Hyun, Jin-Won;Hwang, Eung-Soo;Cha, Chang-Yong;Kook, Yoon-Hoh;Kim, Bum-Joon
    • Journal of Microbiology and Biotechnology
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    • v.17 no.4
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    • pp.701-704
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    • 2007
  • This report describes the full-length sequences of 2HBV clones from a hepatocellular carcinoma (HCC) patient, one with preC mutation (1896A) and the other without preC mutation. The high level of discrepancy in mutation frequency between these 2 strains was observed in the Core (C) region among 4 ORFs. These data support previous results that Korean HBV strains, belonging to genotype C2, are prone to mutations. It is possible that the mutations (BCP and preC mutations) associated with the HBeAg defective production might contribute to the diversity of mutations related to HBV persistence, playing an important role in hepatocarcinogenesis in this patient.

Biochemical Properties of Second Site Mutation of Human Immunodeficiency Virus Integrase

  • Kim, Do-Jin;Oh, You-Take;Lee, Sang-Kwang;Shin, Cha-Gyun
    • BMB Reports
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    • v.32 no.6
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    • pp.599-604
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    • 1999
  • A highly conserved amino acid, glutamic acid (Glu), present at position 152 in the catalytic domain of the human immunodeficiency virus type 1 (HIV-1) integrase (IN) protein has been known to be critical for enzymatic function since substitution of Glu 152 with other residues results in a complete loss of enzymatic activities. In order to better understand the role of Glu 152 as a conserved residue in enzymatic action, intragenic second site mutations have been introduced around residue 152 of a mutant IN (E152A), and their biochemical properties were analyzed in terms of enzymatic activities. Disintegration activities were found to be significantly restored in several second site mutant INs, while integration activities were only recovered weakly. However, endonucleolytic activities were not discovered in all the mutant INs. These findings indicate that the second site mutations can partially restore that catalytic structure of the active site disturbed by the E152A mutation and lead to the regaining of integration and disintegration activities. In addition, it is also suggested that endonucleolytic activity requires a more accurate structure of the catalytic site than that for the integration and disintegration activities.

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