• Title/Summary/Keyword: E7

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Intratypic Variants of HPV-16 E6jE7 Oncogene Isolated from Sexually High-Risk Women in Busan. (부산지역 유흥업소 종사여성으로부터 분리된 HPV16형의 발암유전자(E6/E7) 돌연변이 유형 분석)

  • Min, Sang-Kee;Kim, Sung-Soon;Choi, Byeong-Sun;Jang, Dai-Ho;Lee, Mee-Ok;Choi, Seung-Hwa;Kim, Nam-Ho;Park, Yon-Koung;Jeong, Yeong-A;Kim, Seong-Joon;Bin, Jae-Hun;Park, Ho-Kuk
    • Journal of Life Science
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    • v.19 no.6
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    • pp.765-769
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    • 2009
  • Recent studies have reported that the distribution of HPV-16 sequence variation differs geographically, and more specifically that HPV-16 E6/E7 intratypic variants might carry a high risk for development of ICC (invasive cervical cancer) and CIN (cervical intraepithelial neoplasia) in a given population. To investigate the genetic diversities of HPV-16 E6/E7 oncogene by region, we collected nineteen HPV-16 isolates from sexually high-risk women in Busan, and analyzed the HPV-16 E6/E7 coding regions (nt 34 to 880) with HPV-16 E6/E7 specific PCR amplification. At the nucleotide levet eleven variants of the E6 genes and nine variants of the E7 genes were identified as follows: E6 T178G (n=l1), E6 T178A (n=l), E6 T350G (n=3), E6 A442C (n=2), E6 AI04T, E6 All1G, E6 C116T, E6 G145T, E6 T183G, E6 C335T, E6 G522C and E7 A647G (n=12), E7 A645C, E7 A777C, E7 G663A, E7 T732C, E7 T760C, E7 A775T, E7 T789C and E7 T795G, respectively. At the amino acid levet the isolated HPV-16 E6 and E7 genes showed eleven E6 variants: E6 D25E (n=12), E6 L83V (n=4), E6 E113D (n=2), E6 MIL, E6 Q3R, E6 P5S, E6 Q14H, E6 D25N, E6 127R, E6 H78Y, E6 C140S and three E7 variants: N29S (n=12), L28F, T72S. HPV16 E6 L83V, the dominant variant in the Caucasian population, showed relatively low frequencies in our study population. We elucidated that the dominant HPV-16 E6/E7 variants were HPV-16 E6 D25E (63.2%) and HPV-16 E7 N29S (63.2%), which were phylogenetically included in Asian lineage. Further study is needed to evaluate the risk of cervical cancer related HPV-16 E6/E7 intratypic variants in the Korean population.

Synthesis of Adenosine and Uridine Analogues Containing Conjugated Diene (공액 이중 결합을 갖는 Adenosine과 Uridine 유사체의 합성)

  • Ro, Bong Oh
    • Journal of the Korean Chemical Society
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    • v.45 no.4
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    • pp.312-317
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    • 2001
  • 7'-Aldehyde-nucleosides analogue (2a, 2c) were synthesized from 6-N-benzoyl-2',3'-O-isopropylideneadenosine and uridine. The condensation of 2 with ethoxycarbonylmethylene Wittig reagent produced the adenosine and uridine analogues containing extended conjugated diene and ethoxycarbonyl group, ethyl-1',5',6',7',8'-pentadeoxy-1'-(adenin-9-yl)-$\beta$-D-ribo-nona-5'(E),7'(E)-dienofuranuronate (4b) and ethyl-1'. 5',6',7',8'-pentadeoxy-1'-(uracil-1-yl)-$\beta$-D-ribo-nona-5'(E),7'-(E)-dienofuranuronate (4c). 9-[8',8'-dibromo-5',6',7',8' -tetradeoxy-$\beta$-D-ribo-octa-5'(E),7'(E)-diene]nucleosides (6b, 6c) were also prepared from 2 with similar method.

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Establishment of Transgenic Mouse with the E6 and E7 Genes of Human Papillomavirus Type 16 (인간 Papillomavirus의 E6, E7 유전자를 이용한 Transgenic Mouse의 확립)

  • Hwang, Yong-Il;Lee, Seung-Cheol;Kim, Hyun-Su
    • The Journal of Korean Society of Virology
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    • v.26 no.1
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    • pp.115-120
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    • 1996
  • Human papillomavirus (HPV), especially type 16 and 18, has been closely associated with carcinomas and uterine cevical cancer, recently. From in vitro assays, E6 and E7 genes of HPV16 are closely linked with transformation of cell lines of rodent fibroplasts. However, the transforming activity of E6 and E7 genes of HPV type 16 in vivo has not been fully elucidated. For explaining this mechanism, we prepared a expression system with the promoter of mouse mammary tumorvirus long terminal repeat and E6E7's open reading frames. This expression system was introduced in rodent cell lines, No. 7, 3Y1 and shown normal transforming abilities. And, we produced transgenic mice with E6, E7 expression system. These transgenic mice were confirmed from Southern blot analysis. One male of them was observed enlargement of the testis after 5 months postdelivery.

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Feasibility Study on the Use of Liposomes for Detecting Food-borne Pathogenic Bacteria (식중독 세균 검출에 있어서 리포좀의 이용 가능성)

  • 김명희;김왕준;신원선;손동화;차성관
    • Food Science of Animal Resources
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    • v.23 no.3
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    • pp.278-283
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    • 2003
  • Feasibility tests on using liposomes for detecting food-borne pathogenic bacteria were studied with E. coli 0157:H7 as a model analyte. lmmunoliposomes, whose surface was conjugated with anti-E. coli 0157:H7 IgG and which encapsulated the marker dye, sulforhodamine B, were used for the detection label. Among the feasibility tests, the first test was to use a test-strip on which antibodies to anti-E. coli O157:H7 IgG were immobilized. In this format, immunoliposomes that did not bind to E. coli O157:H7 in sample were captured and then exhibited a visible signal which was inversely related with the number of E. coli O157:H7 in sample. The second test was a direct liposome assay followed by immunomagnetic separation. In this format, immunoliposomes which were bound to E. coli O157:H7 were lysed with detergent and produced a signal which was proportionally related with the number of E. coli O157:H7 in sample. The results from both formats indicate that liposomes can be utilized as a detection label.

Growth Inhibition of Enteropathogenic Escherichia coli $A_2$and Escherichia coli $G_7$ by the Organic Acid Producing Bacteria (유기산 생성균에 의한 병원성 Escherichia Coli $A_2$와 Escherichia Coli $G_7$의 생육억제)

  • 백영진;배형석
    • Microbiology and Biotechnology Letters
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    • v.16 no.2
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    • pp.111-118
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    • 1988
  • The growth inhibition of enteropathogenic Escheriohia coli $A_2$and Escherichia coli G$_7$, causing the diarrhea in piglets, by the organic acid producing bacteria was studied in vitro. The metabolites of the organic acid bacteria, such as lactic acid, acetic acid inhibited the growth of E. coli $A_2$and E. coli G$_7$ in BL medium. The more the organic acid producing bacteria have ability to produce the organic acids, the higher these bacteria excelled the inhibitory efficacy against enteropathogenic E. coli. Among the strains examined, Lactobacillus casei Y and Streptococcus faecium C showed relatively strong growth inhibition against enteropathogenic E. coli.. When the organic acid producing bacteria and the enteropathogenic E. coli were incubated simultaneously in BL medium, bacteriostasis of E. coli was observed when the pH of BL culture was lowered to 5.0, and bacteriocidal effect was observed when the pH became Bess than 4.5, E. coli. $A_2$was more resistant to the organic acid bacteria than E. coli G$_7$.

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Composition of the Aggregation Pheromone Components of Korean Bean Bug and Attractiveness of Different Blends (한국산 톱다리개미허리노린재 집합페로몬 성분의 조성과 배합비율별 유인력)

  • Huh, Hye-Soon;Yun, Ji-Eun;Takashi, Wada;Mizutani, Nobuo;Park, Chung-Gyoo
    • Korean journal of applied entomology
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    • v.47 no.2
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    • pp.141-147
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    • 2008
  • Male adults of bean bug, Riptortus clavatus Thunberg (Heteroptera: Alydidae), release aggregation pheromone (AP) attracting both sexes of adult and nymphs, which its egg parasite, Ooencyrtus nezarae (Hymenoptera: Encyrtidae) exploits the pheromone to find host. The AP consists of three components; (E)-2-hexenyl (Z)-3-hexenoate (E2HZ3H), (E)-2-hexenyl (E)-2-hexenoate (E2HE2H), and tetradecyl isobutyrate (TI). We analyzed composition of the pheromone components of bean bugs from different geo graphical locations of Korea and Japan. The attractiveness of different blends of AP components to R. clavatus was also tested in the fields in Jinju, Korea and in Kumamoto, Japan. Composition ratios (E2HZ3H: E2HE2H:TI) of the AP of Jinju and Iksan populations were 1:1.4:0.2 and 1:0.8:0.2, and those of Tsukuba and Kumamoto populations were 1:2.8:0.2 and 1:1.5:0.1, respectively. In field tests, traps baited with ratio of 1:1:1 (E2HZ3H:E2HE2H:TI=16.7:16.7:16.7mg/rubber septum) and 1:1:0.5(E2HZ3H:E2HE2H:TI= 20:20:10mg/rubber septum) attracted significantly greater number of adult bugs than that of 1:5:1 (E2HZ3H:E2HE2H:TI=7.1:35.7:7.1mg/rubber septum).

Antimicrobial Activity of Flos Syzygii Aromatici Extracts against Mice Infected with Escherichia coli O157:H7 (Escherichia coli O157:H7에 감염된 마우스에 대한 정향 추출물의 항균효과)

  • Lee, Soo-Mi;Son, Song Ee;Lee, Hu Jang
    • Journal of Food Hygiene and Safety
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    • v.32 no.4
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    • pp.336-340
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    • 2017
  • The present study evaluated the antibacterial effect of Flos syzygii Aromatici methanolic extracts (FSAE). In addition, the effectiveness of FSAE against Escherichia coli O157:H7 infection was studied using ICR female mice. At 24 h after incubation of E. coli O157:H7, FSAE at the concentration of 0.269 (p < 0.05), 0.538 (p < 0.001) and 1.075 mg/mL (p < 0.001) significantly inhibited the growth of E. coli O157:H7 compared to the control group. After single challenge with E. coli O157:H7, forty female ICR mice were divided into four experimental groups which were orally administered with saline (control), 0.538 (group 1), 1.075 (group 2) and 2.15 mg/mL (group 3) of FSAE, respectively. On the 3rd day, the number of fecal E. coli O157:H7 in group 2 (p < 0.05) and group 3 (p < 0.01) was significantly decreased compared to that in the control group. On the 7th day post-treatment, the number of fecal E. coli O157:H7 in all FSAE-treated groups was significantly decreased compared to that in the control group (group 1, p < 0.05; group 2 and 3, p < 0.001). According to the results of the present study, administration of FSAE to mice can reduce the severity of E. coli O157:H7 infection. Therefore, the current study suggests that FSAE could be a good candidate for the treatment of enteric infections in domestic animals.

Expression of Human Papillomavirus Type 16, Prototype and Natural Variant E7 Proteins using Baculovirus Expression System

  • Han, Hee-Sung;Kee, Sun-Ho;Hwang, Soon-Bong;Kim, Hyung-Jun;Cho, Kyung-A;Kim, Yoon-Won;Cho, Min-Kee;Chang, Woo-Hyun
    • The Journal of Korean Society of Virology
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    • v.28 no.1
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    • pp.53-62
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    • 1998
  • Human papillomavirus (HPV) 16, E7 proteins derived from the prototype (Bac73) and natural variant (Bac101) E7 open reading frame were produced in Sf9 insect cells. The variant E7 gene occurred naturally by substitution mutation at the position of 88 nucleotide, resulting serine instead of asparagine. Using E7 specific monoclonal antibody (VD6), both E7 proteins were identified in recombinant baculovirus infected SF9 cells. Radiolabelling and immunoprecipitation analysis revealed that both E7 proteins were phosphoproteins. Immunostaining result showed that E7 proteins were mainly localized in the cytoplasm. Nuclear form of E7 proteins was also detected after a sequential fractionation procedure for removing chromatin structure. Considering that the VD6 recognition site in E7 protein is located within 10 amino acid at the N-terminus, this region appears to be blocked by the nuclear component. Western blot analysis revealed that nuclear form was more abundant than cytoplasmic E7 proteins. Time course immunostaining showed that the primary location of E7 protein was the nucleus and exported to the cytoplasm as proteins were accumulated. These events occurred similarly in both Bac73 and Bac101 infected Sf9 cells, suggesting that these two proteins may have similar biological functions.

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Immunotherapeutic Effects of Dendritic Cells Pulsed with a Coden-optimized HPV 16 E6 and E7 Fusion Gene in Vivo and in Vitro

  • Zhou, Zhi-Xiang;Li, Dan;Guan, Shan-Shan;Zhao, Chen;Li, Ze-Lin;Zeng, Yi
    • Asian Pacific Journal of Cancer Prevention
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    • v.16 no.9
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    • pp.3843-3847
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    • 2015
  • Background: Cervical cancer is the second most common cause of cancer related death of women. Persistent HPV infection, especially with high-risk types such as HPV16 and HPV18, has been identified to be the primary cause of cervical cancer. E6 and E7 are the major oncoproteins of high-risk HPVs, which are expressed exclusively in HPV infected tissues, and thereby represent ideal therapeutic targets for immunotherapy of cervical cancer. Materials and Methods: In this work, we used recombinant adenovirus expressing coden-optimized HPV16 E6 and E7 fusion protein (Ad-ofE6E7) to prime dendritic cells (DC-ofE6E7), to investigate the ability of primed DC vaccine in eliciting antitumor immunity in vitro and vivo. Results: Our results indicated that DC-ofE6E7 vaccine co-culturing with splenocytes could strongly induce a tumor-specific cytotoxic T lymphocyte (CTL) response and kill the TC-1 cells effectively in vitro. Moreover, DC-ofE6E7 vaccine induced protective immunity against the challenge of TC-1 cancer cells in vivo. Conclusions: The results suggested that the HPV16 ofE6E7 primed DC vaccine has potential application for cervical cancer immunotherapy.

A Novel Mutant of Human Papillomavirus Type 18 E6E7 Fusion Gene and its Transforming Activity

  • Zhou, Zhi-Xiang;Zhao, Chen;Li, Qian-Qian;Zeng, Yi
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.17
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    • pp.7395-7399
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    • 2014
  • Background: Persistent human papillomavirus (HPV) infection, especially with high-risk types such as HPV16 and HPV18, has been identified as the primary cause of cervical cancer. E6 and E7 are the major onco-proteins of high-risk HPVs, which are consistently expressed in HPV infected tissues but absent in normal tissues and represent ideal therapeutic targets for immunotherapy of cervical cancer. Materials and Methods: In this study, the optimized fusion gene HPV18 E6E7 (HPV18 ofE6E7) was constructed according to genetic codon usage for human genes. At the same time, for safety future clinical application, a mutant of HPV18 ofE6E7 fusion gene was generated by site-directed mutagenesis at L52G for the E6 protein and C98G for the E7 protein. Results: HPV18-E6E7 mutant (HPV18 ofmE6E7) constructed in this work not only lost the transformation capability for NIH 3T3 cells and tumorigenicity in BALB/c nude mice, but also maintained very good stability and antigenicity. Conclusion: These results suggest that the mutant should undergo further study for application as a safe antigenspecific therapeutic vaccine for HPV18-associated tumors.