• Title/Summary/Keyword: env gene

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Korean Red Ginseng slows coreceptor switch in HIV-1 infected patients

  • Young-Keol Cho;Jung-Eun Kim;Jinny Lee
    • Journal of Ginseng Research
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    • v.47 no.1
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    • pp.117-122
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    • 2023
  • Background: Human immunodeficiency virus-1 (HIV-1) that binds to the coreceptor CCR5 (R5 viruses) can evolve into viruses that bind to the coreceptor CXCR4 (X4 viruses), with high viral replication rates governing this coreceptor switch. Korean Red Ginseng (KRG) treatment of HIV-1 infected patients has been found to slow the depletion of CD4+ T cells. This study assessed whether the KRG-associated slow depletion of CD4+ T cells was associated with coreceptor switching. Methods: This study included 146 HIV-1-infected patients naïve to antiretroviral therapy (ART) and seven patients receiving ART. A total of 540 blood samples were obtained from these patients over 122 ± 129 months. Their env genes were amplified by nested PCR or RT-PCR and subjected to direct sequencing. Tropism was determined with a 10% false positive rate (FPR) cutoff. Results: Of the 146 patients naïve to ART, 102 were KRG-naïve, and 44 had been treated with KRG. Evaluation of initial samples showed that coreceptor switch had occurred in 19 patients, later occurring in 38 additional patients. There was a significant correlation between the amount of KRG and FPR. Based on initial samples, the R5 maintenance period was extended 2.35-fold, with the coreceptor switch being delayed 2.42-fold in KRG-treated compared with KRG-naïve patients. The coreceptor switch occurred in 85% of a homogeneous cohort. The proportion of patients who maintained R5 for ≥10 years was significantly higher in long-term slow progressors than in typical progressors. Conclusion: KRG therapy extends R5 maintenance period by increasing FPR, thereby slowing the coreceptor switch.

Molecular Phylogenetic Analysis of HIV -1 vif Gene from Korean Isolates

  • Park, Chan-Seung;Kim, Mi-Sook;Lee, Sung-Duk;Kim, Sung-Soo;Lee, Keon-Myung;Lee, Chan-Hee
    • Journal of Microbiology
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    • v.44 no.6
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    • pp.655-659
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    • 2006
  • Phylogenetic studies of nef, pol, and env gene sequences of HIV-1 isolated from Koreans suggested the presence of a Korean clade in which Korean sequences are clustered to the exclusion of foreign sequences. We attempted to identify and characterize the Korean clade using all vif gene sequences isolated from Koreans registered in the NCBI GenBank database (n = 233). Most (77 %) of the Korean isolates belonged to the Korean clade as a large subcluster in subtype B, designated the Korean clade subtype B ($K_{C}B$). $K_{C}B$ sequences were relatively homogenous compared to Korean subtype B sequences that did not belong to the $K_{C}B$ (non-Korean clade subtype B; $NK_{C}B$). Comparison of amino acid frequencies of $K_{C}B$ and $NK_{C}B$ sequences revealed several positions where the amino acid frequencies were significantly different. These amino acid residues were critical in separating $K_{C}B$ from $NK_{C}B$ or from foreign sequences, since substitution of these amino acids in $K_{C}B$ with the $NK_{C}B$ amino acids relocated the $K_{C}B$ sequences to $NK_{C}B$, and vice versa. Further analyses of $K_{C}B$ will help us to understand the origin and evolutionary history of $K_{C}B$.

Investigation of genomic integration of GX-12, a new anti-HIV DNA vaccine, into host cellular DNA following intramuscular injection in rats

  • Kang, Kyung-Koo;Park, Min-Seul;Ahn, Jun-Kook;Baik, Dae-Hyun;Lee, Dong-Sup;Park, Jae-Hun;Ahn, Byoung-Ok;Kim, Won-Bae
    • Proceedings of the Korean Society of Toxicology Conference
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    • 2002.11b
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    • pp.199-200
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    • 2002
  • GX-12 is a naked DNA vaccine developed by research team of Dong-A Pharmaceutical Company, Green Cross Company and Genexine for the treatment of HIV infection. It consists of four separate plasmids (pGX10-GE HX, pGX10-dpol JR, pGX10-VN/TV JR, pGX10-hIL-12m), which were constructed by inserting the HIV-1 gag-env, pol, regulatory genes and a human IL-12 mutant gene into pGX10 plasmid vectors.(omitted)

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Diagnosis of Bovine Leukemia Virus (BLV) infection using PCR and ELISA techniques in Holstein dairy cattle (홀스타인종 젖소에 있어서 PCR과 ELISA기법을 이용한 BLV 감염진단)

  • Jeong, Hang-Jin;Yu, Seong-Lan;Lee, Jun-Heon;Do, Chang-Hee;Shu, Guk-Hyun;Ryoo, Seung-Heui;Sang, Byung-Chan
    • Korean Journal of Agricultural Science
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    • v.38 no.1
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    • pp.45-50
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    • 2011
  • This study was conducted to investigate the farm situation about bovine leukemia virus(BLV) infection that greatly influence productivity in dairy cattle and compare the accuracy of diagnosis for BLV infection between PCR and ELISA techniques. Blood samples of 193 heads from 5 herds in Chungnam and Chungbuk area were used to analyze BLV gene and serum, and the results were obtained as follows. The amplified BLV gene in dairy cattle by PCR technique resulted in 226 bp, 596 bp and 434 bp, respectively, for gag, pol and env, which were well amplified. The infection rates of BLV virus diagnosed by PCR and ELISA techniques ranged from 80.55 to 100% and from 22.22 to 86.95%, respectively, and the infection rates among 5 herds were significantly different in both methods (P<0.05). Further, the average infection rates of 5 herds were 87.05 and 63.21%, respectively, for PCR and ELISA techniques. Kappa statistics for examining consistency of diagnosis by PCR and ELISA techniques showed 0.246, which represents low consistency. Consequently, PCR based BLV technique was considered as a corrective measure for diagnosis of BLV infection in Holstein dairy cattle.

Transformation of Pleurotus sajor-caju by Complementation of PABA Requiring Mutant (여름 느타리버섯에서 PABA 변이주의 Complementation에 의한 형질전환)

  • Byun, Myung-Ok;Chung, Jong-Chun;You, Chang-Hyun;Cha, Dong-Yeul;Lee, Du-Hyung
    • The Korean Journal of Mycology
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    • v.25 no.3 s.82
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    • pp.233-237
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    • 1997
  • A PABA auxotroph of Pleurotus sajor-caju were transformed to prototrophy by using a plasmid containing pab 1 gene from Coprinus. The efficiencies of transformation of Pleurotus sajor-caju was five transformants per ${\mu}g$ of plasmid DNA. Southern blot analysis of DNA extracted from transformants demonstrated that plasmid DNA was integrated into the chromosomal DNA in multiple tandem copies. Progenies of heterokaryons between transformants of PABA and other auxotropic strains produced pab-progeny, which indicated that integration occurred at a site(s) other than the resident pab biosynthetic gene.

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Inhibition of Porcine Endogenous Retrovirus Expression by RNA Interference (RNA 간섭을 통한 Porcine Endogenous Retrovirus의 발현 억제)

  • Lee, Hyun-A;Koo, Bon-Chul;Kwon, Mo-Sun;Kim, Te-Oan
    • Reproductive and Developmental Biology
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    • v.30 no.3
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    • pp.181-187
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    • 2006
  • In recent years the number of patients waiting for organ transplantation has greatly outpaced the supply of human organs available, which leads to a renewed interest in pig-to-human xenotransplantation as an alternative. However, one of the biggest barriers in the xenotransplantation is presence of porcine endogenous retroviruses(PERV) that can infect human cells. In this study, to present a possible solution for this problem we tried to inhibit expression of PERVs using shRNAs(short hairpin RNA) at the level of RNA synthesis and virus release. The shRNA targeting the sequence of PERV A, B type was cloned into pSIREN-RetroQ vector under the control of polymerase-III U6-RNA gene promoter. Quantitative real-time PCR was performed to detect my alterations in mRNA production of PERV A, B targeted by the shRNA in each done. Depending on the target sequence of the shRNA, the transcription of PERV was decreased to as much as 4% and the number of progeny viruses was reduced to less than 1/200,000. Transgenic pigs producing such shRNAs may result in a highly reduced PERV expression in cells and organs, which is a prerequisite for safe xenotransplantations.

OmpR Is Essential for Growth and Expression of Virulence-related Genes in the Fish Pathogen Edwardsiella piscicida (어류 병원체 Edwardsiella piscicida의 OmpR은 생육과 병원성과 관련된 유전자의 발현에 필수적)

  • Ray, Durga;Kim, Yeon Ha;Choe, unjeong;Kang, Ho Young
    • Journal of Life Science
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    • v.31 no.1
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    • pp.28-36
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    • 2021
  • Edwardsiella piscicida is a significant cause of hemorrhagic septicemia in fish and gastrointestinal infections in humans. Survival bacteria require specialized mechanisms to adapt to environmental fluctuations. Hence, to understand the mechanism through which E. piscicida senses and responds to environmental osmolarity changes, we determined the protein expression profile and physiological properties under various salinity conditions in this study. The OmpR protein is a part of the Env-ZOmpR two-component system that has been implicated in sensing salt stress in bacteria. However, the physiological role played by this protein in E. piscicida remains to be elucidated. Therefore, in this work, the function of the OmpR protein in response to salt stress was investigated. Phenotypic analysis revealed that, in the mutant, three of the biochemical phenotypes were different from the wild type, including, citrate utilization, hydrogen sulfide, and indole production. Introduction of the plasmid containing the entire ompR gene to the mutant strain returned it to its parental phenotype. The retarded growth rate also partially recovered. Furthermore, in our studies, OmpR was not found to be related to cell motility. Taken together, our results from the mutational analysis, the growth assay, MALDI-TOF MS, qRT-PCR, and the phenotype studies suggest that the OmpR of E. piscicida is implicated in osmoregulation, growth, expression of porins (ETAE_1826), virulence-related genes (EseC, EseD and EvpC), and certain genes of unknown function (ETAE_1540 and ETAE_2706).

Detection of BLV Proviral DNA in Korean Native Goats Experimentally Infected with Bovine Leukemia Virus by Polymerase Chain Reaction (소백혈병 바이러스 (Bovine Leukemia Virus)에 감염된 한국 재래산양에서 PCR기법을 이용한 BLV 유전자 검출)

  • Jun, Moo-Hyung;Chang, Kyung-Soo;Cho, Young-Sung;Park, Jong-Hyeon;An, Soo-Hwan
    • The Journal of Korean Society of Virology
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    • v.27 no.2
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    • pp.217-225
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    • 1997
  • PCR amplication using the primers for gag, pol and env genes in BLV (bovine leukemia virus) proviral DNA and syncytium assay were carried out for the Korean native goats experimentally infected with bovine leukemia virus to investigate pathogenesis of BLV in the goats, and to establish a model animal for BLV infection. The oligonucleotide primers used in PCR revealed very high specificity. The minimal amount of FLK-BLV cellular chromosomal DNA to detect the integrated BLV proviral DNA was 10 ng. The peripheral blood lymphocytes from the goat infected with BLV were examined at regular intervals by PCR amplification and syncytium assay. Pol or gag genes were detected in none of three infected goats at the 1st week post-infection (p.i.). At the 4th week p.i., one of three goats showed the amplified gag gene. Thereafter detection rates for the genes were increased, indicating that the BLV proviral genes were integrated in all of the lymphocytes from three goats, at the 16th weeks p.i., when it was evident in syncytium assay that the lymphocytes from all of three goats were infested with infective BLV. Investigating the tissues from the necropsied goats at the 8th month p.i., the amplified BLV proviral genes and infective BLV were detected in all of the peripheral lymphocytes from three infected-goats. Among various tissues examined, the amplified BLV proviral genes were observed in spleen and superficial cervical, mandibular and retropharyngeal lymph nodes, and the infective BLV, in superficial cervical and mandibular lymph nodes. It was assumed that the Korean native goat was quite susceptible to BLV infection, indicating that the goat could be a good model animal for BLV.

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Complete Sequences of HIV-1 in a Korean Long-term Nonprogressor with HIV-1 Infection (장기간 진행하지 않는 인면역결핍바이러스(Human Immunodeficiency Virus, HIV)-1 감염자로부터 분리한 HIV-1의 전체 염기서열 결정)

  • Cho, Young-Keol;Lee, Hee-Jung;Desrosiers, Ronald C.
    • The Journal of Korean Society of Virology
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    • v.29 no.2
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    • pp.107-118
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    • 1999
  • To characterize the molecular nature of human immunodeficiency virus (HIV)-1, we determined the full-length HIV-1 sequences from cultured peripheral blood mononuclear cells (PBMC) of a Korean long-term nonprogressor (LTNP). Without antiretroviral therapy, the individual has maintained CD4+ T counts over $500/{\mu}l$ from 1989 to 1999. Plasma viral RNA copy was 992 U/ml in 1998. Culture supernatant showed positive from culture days 9. A series of 9 overlapping PCR products were amplified from cultured PBMC and cloned About 9.2 kb from R of 5' LTR to R of 3' LTR was determined by automated sequencing. The G-to-A hypermutations were shown throughout the entire region. As a result of G to A hypermutations, premature stop codon was found in integrase coding region. Though there was no recombination between subtypes over all genomes, TATA box in both LTRs was TAAAA which is detected in subtype E instead of TATAA in subtype B. And, there were nucleotide GC insertion between $NF-{\kappa}B$ I and Sp1 III, and duplication of $TCF-1{\alpha}$ in LTR. We could not find any deletion of amino acid in Nef, Gag, Pol and Env gene. This study is the first report on molecular nature of full genomes of HIV-1 isolated in Korea.

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The frequency of defective genes in vif and vpr genes in 20 hemophiliacs is associated with Korean Red Ginseng and highly active antiretroviral therapy: the impact of lethal mutations in vif and vpr genes on HIV-1 evolution

  • Cho, Young Keol;Kim, Jung-Eun
    • Journal of Ginseng Research
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    • v.45 no.1
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    • pp.149-155
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    • 2021
  • Background: We have reported that internal deletions in the nef, gag, and pol genes in HIV-1-infected patients are induced in those treated with Korean Red Ginseng (KRG). KRG delays the development of resistance mutations to antiretroviral drugs. Methods: The vif-vpr genes over 26 years in 20 hemophiliacs infected with HIV-1 from a single source were sequenced to investigate whether vif-vpr genes were affected by KRG and KRG plus highly active antiretroviral therapy (ART) (hereafter called GCT) and compared the results with our previous data. Results: A significantly higher number of in-frame small deletions were found in the vif-vpr genes of KRG-treated patients than at the baseline, in control patients, and in ART-alone patients (p < 0.001). These were significantly reduced in GCT patients (p < 0.05). In contrast, sequences harboring a premature stop codon (SC) were more significant in GCT patients (10.1%) than in KRG-alone patients, control (p < 0.01), and ART-alone patients (p = 0.078 for peripheral blood mononuclear cells). The proportion of SC in Vpr was similar to that in Vif, whereas the proportion of sequences revealing SC in the env-nef genes was significantly lower than that in the pol-vif-vpr genes (p < 0.01). The genetic distance was 1.8 times higher in the sequences harboring SC than in the sequences without SC (p < 0.001). Q135P in the vif gene is significantly associated with rapid progression to AIDS (p < 0.01). Conclusion: Our data show that KRG might induce sD in the vif-vpr genes and that vif-vpr genes are similarly affected by lethal mutations.