• Title/Summary/Keyword: Glycoprotein(G) gene

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Sequencing and Baculovirus-Based Expression of the Glycoprotein B2 Gene of HSV-2 (G)

  • Uh, Hong-Sun;Park, Jong-Kuk;Kang, Hyun;Kim, Soo-Young;Lee, Hyung-Hoan
    • Journal of Microbiology and Biotechnology
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    • v.11 no.3
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    • pp.482-490
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    • 2001
  • The gene for glycoprotein B (gB2) of HSV-2-strain G was subcloned, sequenced, recombinated into the lacZ-HcNPV, expressed in insect cells, and compared with the homologous gene of other HSV-2 strains. The ORF of the gB2 gene was 2,715 bp. The overall nucleotide sequence homology of te gB2 gene compared ith that of the two previously reported HSV-2 strains appeared to be over 98%. A recombinant virus named Baculo-gB2 protein in insect cells. The recombination was confirmed by a PCR and the expression was demonstrated by radio immunoprecipitation. Insect cells infected with the Baculo-gB2 virus synthesized and processed gB2 with approximately 120 kDa in the cells, and then secreted it into the culture media, where it reacted with a nomoclonal antibody to gB2. The gB2 polypeptide contained two main hydrophobic regions (a signal sequence from 1 to 23 amino acid residues, and a membrane anchor sequence from aa 745 to 798), eight N-glycosylation sites evenly distributed, and was rich in alanine (11.2%). Antibodies to this recombinant protein that were raised in mice recognized the viral gB2 and neutralized the infectivity of the HSV-2 in vitro. There results show that the gB2 protein was successfully porduced in insect cells and could be used to raise a protective neutralizing antibody. Accordingly, this particular recombinant protein may be useful in the development of a subunit vaccine.

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A Replication-Competent Retroviral Vector Expressing the HERV-W Envelope Glycoprotein is a Potential Tool for Cancer Gene Therapy

  • Byoung Kwon Kang;Yong-Tae Jung
    • Journal of Microbiology and Biotechnology
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    • v.34 no.2
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    • pp.280-288
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    • 2024
  • The fusogenic membrane glycoprotein (FMG) derived from the human endogenous retrovirus-W (HERV-W) exhibits fusogenic properties, making it a promising candidate for cancer gene therapy. When cells are transfected with HERV-W FMG, they can fuse with neighboring cells expressing the receptor, resulting in the formation of syncytia. These syncytia eventually undergo cell death within a few days. In addition, it has been observed that an HERV-W env mutant, which is truncated after amino acid 483, displays increased fusogenicity compared to the wild-type HERV-W env. In this study, we observed syncytium formation upon transfection of HeLa and TE671 human cancer cells with plasmids containing the HERV-W 483 gene. To explore the potential of a semi-replication-competent retroviral (s-RCR) vector encoding HERV-W 483 for FMG-mediated cancer gene therapy, we developed two replication-defective retroviral vectors: a gag-pol vector encoding HERV-W 483 (MoMLV-HERV-W 483) and an env vector encoding VSV-G (pCLXSN-VSV-G-EGFP). When MoMLV-HERV-W 483 and pCLXSN-VSV-G-EGFP were co-transfected into HEK293T cells to produce the s-RCR vector, gradual syncytium formation was observed. However, the titers of the s-RCR virus remained consistently low. To enhance gene transfer efficiency, we constructed an RCR vector encoding HERV-W 483 (MoMLV-10A1-HERV-W 483), which demonstrated replication ability in HEK293T cells. Infection of A549 and HT1080 human cancer cell lines with this RCR vector induced syncytium formation and subsequent cell death. Consequently, both the s-RCR vector and RCR encoding HERV-W 483 hold promise as valuable tools for cancer gene therapy.

Genetic relationship of the VHSV(Viral Hemorrhagic Septicemia Virus)isolated from cultured olive flounder, Paralichthys olivaceus in Korea (우리나라 양식 넙치, Paralichthys olivaceus에서 분리된 VHSV(Viral Hemorrhagic Septicemia Virus)의 유전학적 검토)

  • Kim, Su-Mi;Lee, Jae-Il;Hong, Mi-Ju;Park, Heon-Sik;Park, Su-Il
    • Journal of fish pathology
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    • v.16 no.1
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    • pp.1-12
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    • 2003
  • RT-PCR method was applied to detect and clone the nucleocapsid protein (N) gene and glycoprotein (G) gene for sequencing 5 Korean VHSV isolates from cultured olive flounder, Paralichthys olivaceus. Phylogenetic analysis was performed to investigate their relationship with the VHSV strains described previously and isolated from different geographical area. Generally, VHSV strains were separated phylogenetically according to the major geographical area of isolation: Genogroup I (American type), Genogroup Il (British Isles) and Genogroup ill (European type). This study revealed that all 5 Korean VHSV isolates were belonged to Genogroup I and closely related to Japanese Obama25 type.

Pharmacokinetic Characteristics of Levosulpiride in Relation to the Genetic Polymorphism of MDR1: From Knockout Mouse to Human

  • Cho, Hea-Young;Lee, Yong-Bok
    • Journal of Pharmaceutical Investigation
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    • v.37 no.5
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    • pp.297-303
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    • 2007
  • The purposes of this study were to clarify the involvement of P-glycoprotein (P-gp) in the efflux of levosulpiride in knockout mice that lack the mdr1a1b gene and to evaluate the relationship between the genetic polymorphisms in MDR1 gene (exon 21) and levosulpiride disposition in healthy Korean subjects. After oral administration ($10\;{\mu}g/g$) of levosulpiride to mdr1a/1b(-/-) and wild-type mice, plasma and brain samples were obtained at 45 min. We also investigated the genotype for MDR1 (exon 21) gene in humans using a polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method. A single oral dose of 25 mg levosulpiride was administered to 58 healthy subjects, who were based on the MDR1 genotype for the G2677T SNP. Blood samples were taken up to 36 hr after dosing. The concentrations of levosulpiride in mouse plasma and brain were statistically significant difference between the two animal groups (P<0.05). In addition, the average brain-to-plasma concentration ratio (Kp) of levosulpiride was 3.4-fold (P<0.01) higher in the mdr1a/1b(-/-) mice compared with the wild-type mice. We also found that the values of $AUC_{0-{\infty}$, partial AUC ($AUC_{0-4h}$) and $C_{max}$ were significantly different between homozygous 2677TT subjects and the subjects with at least one wild-type allele (GG and GT subjects, P=0.012 for $AUC_{0-{\infty}$; P=0.008 for $AUC_{0-4h}$; P=0.038 for $C_{max}$). The results confirm that levosulpiride is a P-gp substrate in vivo, and clearly demonstrate the effect of SNP 2677G>T in exon 21 of the MDR1 gene on levosulpiride disposition.

Cloning of the non-virion (NV) of a Korean Isolate of Infectious Hematopoietic Necrosis and Identification of the Role of the NV in IHNV Replication (한국에서 분리된 전염성 조혈괴저 바이러스의 non-virion (NV) 단백질의 유전자 클로닝 및 바이러스 증식에서의 역할)

  • 문창훈;조화자;윤원준;박정재;박정민;김현주;도정완;이주양;임채렬
    • Korean Journal of Microbiology
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    • v.36 no.2
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    • pp.103-108
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    • 2000
  • We have cloned and analyzed cDNA coding for non-virion (NV) protein of the m V - P R T The NV gene contained 336 bp open readmg frame and encoded a protein of 11 1 amino acids with a molecular weight of 13.2 kDa. The deduced amino acid sequence of NV of IHNVPRT was found to be 90-95% identical to those of foreign isolates of IHNV. These results indicate that NV gene of the MNV is highly conserved among &ifferent strains of THNV Northern blot analyses revealed that the levels of NV gene expression were strongly elevated after 20 h post-infection. In order to identify the role of NV in the replication of MNV in fish cells, IHNVinfected cells were treated with antisense oligonucleotides. While IHNV-PRT exposed to glycoprotein (G) antisense oligonucleotide showed severely reduced growth, the growth of virus exposed to NV antisense oligonucleotide was not affected by NV antisense oligonucleotide, which suggests that NV is not essential for replication of IHNV in fish cells.

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Retrovirus Vector-Mediated Inductional Expression of the Human Lactadherin Gene in Mouse Mammary Epithelial Cells (Mouse Mammary Epithelial Cell에서 Retrovirus Vector를 이용한 Human Lactadherin 유전자의 유도적 발현)

  • 권모선;구본철;정병현;염행철;박창식;김태완
    • Korean Journal of Animal Reproduction
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    • v.27 no.1
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    • pp.15-23
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    • 2003
  • Lactadherin (formerly known as BA46), a major glycoprotein of the human milk fat globule membrane, is abundant in human breast milk and breast carcinomas and may prevent symptomatic rotavirus infections. In this study, under the control of tissue specific and hormonal inducible mouse whey acidic protein (WAP) promote., the expression pattern of lactadherin (Ltd) in lactogenic hormone-dependent mouse mammary epithelial cell line HC11 were tested. pLNWLtd construct containing 2.4 kilobases of the WAP promoter and 1.5 kilobases of human lactadherin gene was stably transfered into HC11 cells using retroviral vector system. Integration and expression level of the transgene was estimated using PCR and RT-PCR, respectively. Prominent induction of Ltd gene under the WAP promoter was accomplished in the presence of insulin, hydrocortisone and prolactin. Compared to the control (cells cultured with insulin alone), however we observed that the WAP promoter was leaky. These data indicate that luther studies are needed in finding an appropriate promoter other than WAP promoter because of its leakiness.

Monitoring of VHS and RSIVD in cultured Paralichthys olivaceus of Jeju in 2014 (2014년 제주도 양식 넙치, Paralichthys olivaceus에 대한 VHS 및 RSIVD 모니터링)

  • PARK, Hyun-Kyung;KIM, Seung-Min;LEE, Da-Won;JUN, Lyu-Jin;JEONG, Joon-Bum
    • Journal of Fisheries and Marine Sciences Education
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    • v.27 no.3
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    • pp.879-889
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    • 2015
  • The outbreak of viral diseases caused by viral haemorrhagic septicaemia virus (VHSV) and red seabream iridovirus (RSIV) have been reported in cultured olive flounder, Paralichthys olivaceus. VHSV has been a serious viral disease that infects the olive flounders in South Korea. Clinical signs of VHSV infection are skin darkening, abdominal distension and haemorrhages. Outbreaks of fish iridoviral disease was first reported from red seabream, Pagrus major farms in Japan. Recently, iridovirus infection have occurred frequently from olive flounder farms in South Korea. In this study, disease surveillance was performed to monitor the prevalence of VHSV and RSIV in olive flounder in 2014. The samples were collected from 60 different olive flounder farms in Jeju from April, May, September, November and December in 2014. RT-PCR (VHSV) or PCR (RSIV) results showed that VHSV were detected in 5 farms, but RSIV has not been detected in any farms. The migration of olive flounder was restricted for the quarantine in 5 farms of VHS outbreak. The nucleocapsid protein (N) gene and glycoprotein (G) gene sequences of the 5 Korean VHSV isolates were successfully amplified and sequenced. Phylogenetic analysis was performed using the VHSV sequences reported here together comparison with the nucleotide sequences available from the GenBank database. Phylogenetic analysis indicated that most of Korea VHSV belong to the genotype IVa and closely related to the strains from Japan and China.

Detection of viral hemorrhagic septicemia virus (VHSV) in wild marine fishes in the coastal region of Korea (우리 나라 연근해 자연산 해수 어종에서의 Viral Hemorrhagic Septicemia Virus (VHSV)의 검출)

  • Kim, Su-Mi;Park, Su-Il
    • Journal of fish pathology
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    • v.17 no.1
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    • pp.1-10
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    • 2004
  • In order to analyse the detection of viral hemorrhagic septicemia virus (VHSV) in marine environment surrounding coastal region of Eastern and Southern sea of Korea, the pools of each organ sample of three fish were taken for virus assay from February to May in 2003. The samples comprised 42, taken from 9 species of marine fishes. The VHSV was detected from chub mackerel Scomber japonicus and striped mullet Mugil cephalus in epithelioma papulosum cyprini (EPC) cells. The identity of the virus was confirmed a reverse transcriptase-polymerase chain reaction (RT-PCR). VHSV has previously been reported from chub mackerel, but not from striped mullet. The new isolates was classified as a member of genogroup I (American type) of VHSV and was closely related to the VHSV KVHS'01-l based on comparisons of the partial nucleotide sequence of the glycoprotein (G) gene.

Genetic positioning of Korean viral hemorrhagic septicemia virus (VHSV) from cultured and wild marine fishes

  • Kim, Wi-Sik;Jung, Sung-Ju;Kim, Jong-Oh;Kim, Du-Woon;Kim, Jeong-Ho;Oh, Myung-Joo
    • Journal of fish pathology
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    • v.24 no.1
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    • pp.1-9
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    • 2011
  • Viral haemorrhagic septicaemia virus (VHSV) is an epidemic virus in olive flounder Paralichthys olivaceus farms in Korea, since the virus have first isolated in 2001. In the present study, partial glycoprotein (G) gene nucleotide sequences of seven Korean VHSV from cultured olive flounder and wild marine fishes in coastal areas of Korea were analyzed to evaluate their genetic relatedness to worldwide isolates. Phylogenetically, all Korean VHSV formed only one minor cluster including Japanese isolates, in genotype IVa, while the North America isolates formed a different minor cluster in genotype IVa. These results suggest that Korean VHSV could be an indigenous virus in Korean and Japanese coastal areas, but have not been introduced from North America.