• Title/Summary/Keyword: virus resistant

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Production of a transgenic pig expressing 3D8 single chain variable fragment (scFv) and its evaluation of PRRS resistant (3D8 scFv 형질전환 돼지 개발 및 PRRS 저항성 평가)

  • Lee, Hwi-Cheul;Lee, Gunsup;Kim, Ji-Yoon;Yang, Hyeon;Lee, Bo Ram;Park, Mi-Ryung;Hwang, In-Sul;Lee, Poongyeon;Byun, Sung-June;Kim, Won-Il;Oh, Keon Bong
    • Korean Journal of Veterinary Service
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    • v.43 no.4
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    • pp.227-236
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    • 2020
  • In this study, we have developed 3D8 scFv transgenic pig (TG) by microinjection of fertilized one-cell pig zygotes (2.17%). The effect of 3D8 scFv TG on porcine reproductive and respiratory syndrome virus (PRRSV) resistance were evaluated through PRRSV VR2332 (1×103 TCID50/mL) challenge and transmission experiments. As a result, the average daily weight gain (ADWG) of TG increased compared to the wild type pigs (WT) in PRRSV challenge groups and the serum viremia levels of the TG was significantly lower than of WT on the 7 day and 21 day after infection, meaning that the viral shedding was suppressed by 3D8 scFv expression. These results suggest that the expression of 3D8 scFv in pig could suppress spreading of infected virus to pigs sharing a room.

Histological Observations and Comparison of the Resistance to Polyhedrosis Viruses in Various Varieties of the Silkworm, Bombyx mori L., fed on Artificial Diet (가잠의 인공사료육에 있어서 다각체 바이러스에 대한 품종별 저항성 및 조직학적 관찰)

  • Gang, Seok-U;Im, Jong-Seong;Son, Hae-Ryong
    • Journal of Sericultural and Entomological Science
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    • v.27 no.2
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    • pp.20-27
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    • 1985
  • This study was carried out to investigate the histological changes after the infection of nuclear and cytoplasmic polyhedrosis viruses(NPV, CPV) and the resistance to the viruses in various varieties of the silkworm fed on artificial diet. The results obtained were as follows; Among four varieties of silkworm tested, Jam 107$\times$Jam 108 was more resistant than the other varieties tested and Jam 119$\times$Jam 120 was the most resistant to CPV. In case of peroral infection with NPV, Jam 107$\times$Jam 108 showed lower mortality than the remained varieties in low concentration (104/ml). However, all varieties showed high mortality as the concentration of viruses was increased. With infection of CPV, the varieties showed high mortality at the concentration of 107 and 108/ml, while Jam 119$\times$Jam 120 showed the lowest mortality at virus concentration of 104/ml. The fat bodies, epidermal cells and tracheal epithelial cells showed high susceptibility to NPV to break the cells completely and liberate the debris to the body cavity. The CPV infected only the cylindrical cells of mid-gut and formed polyhedrons. In some cells, CPV was liberated to gastral cavity. In the electrophoretic pattern of hemolymph protein of silkworm larvae infected with NPV, bands were dimmed and disappeared as symptom aggravated after infection. Electrophoretic pattern of homolymph proteins of silkworm larvae infected with CPV showed no numerical difference at the later stage of infection, and one or two bands was observed along with lowering the concentrations.

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Identification of Introduced Gene and Its Expression and Gene Stability Assessment for Event Selection of Genetically Modified Plant toward Approval: Cucumber Mosaic Virus Resistant Hot Pepper (상업용 유전자 변형작물 이벤트 선발을 위한 도입유전자 확인, 발현 및 세대간 안정성 평가 : 오이모자이크바이러스 저항성 GM 고추)

  • Kang, Seung-Won;Han, Bal-Kum;Lee, Tae-Ho;Kim, Eun-Ji;Lee, Gung-Pyo
    • Horticultural Science & Technology
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    • v.30 no.2
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    • pp.192-200
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    • 2012
  • For the development of genetically modified plants, it is important to verify various factors which potentially affect the risk assessment as well as to establish an experimental program to produce scientific and reliable data. However, it is a time and cost consuming process to develop GM plants as well as to prepare scientific and convincible data for government's approval. Therefore, using the transgenic hot pepper tolerant to a new CMV pathotype, we attempted to suggest few methodological procedures, such as probe saturation for southern blot analysis and RT-PCR and ELISA for expression analysis, for identification and stability evaluation of inserted gene in genetically modified plant which are required for submission for approval. Ten partially overlapped probes covering full length of inserted gene were produced. We could identify that the inserted gene was stacked as a single copy as well as no partial element existed. Also, we could identify the stability of the inserted gene stacked in hot pepper using probe saturation. In the expression analysis with RT-PCR and ELISA, we also could provide the stable expression of transcript and proteins in leaves and placenta and pericarp of fruits of the CMV-resistant hot pepper.

Regeneration of Fertile Transgenic Rice Plane from a Korean Cultivar, Nakdongbyeo (한국 재배종 낙동벼에서 임성 형질전환식물체의 재분화)

  • Soo In LEE;Hyun Jin CHUN;Chae Oh LIM;Jeong Dong BAHK;Moo Je CHO
    • Korean Journal of Plant Tissue Culture
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    • v.22 no.3
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    • pp.175-182
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    • 1995
  • Rice is one of the most successful monocot in regenerating fertile and genetically stable transgenic plants. However there is no report of a rice line developed in Korea that can be used for regeneration of fertile and genetically stable transformants. In this paper we first demonstrate that a Korean variety Nakdongbyeo, is suitable to obtain transgenic rice plants. Protoplasts from embryogenic suspension cultures were co-transformed with HPT (hygromycin phosphotransferase) and GUS ($\beta$-glucuronidase) genes in separate plasmids in the presence of PEG (polyethylene glycol). In 5 independent experiment, the average frequency of calli showing hygromycin resistance were 1.73%. Plantlets were regenerated from the Hy $g^{R}$ calli. The average efficiency of plantlet regeneration was apprbximately 27%. Based on the GUS activities of hygromycin resistant calli, ca.35% of the resistant calli carried active GUS genes. The R0 transgenic plantlets were grown to maturity and Rl seeds were obtained. By examining the in siぉ activity of GUS in Rl seeds and seedlings, we confirmed that the GUS transgene driven by a CaMV 35S (cauliflower mosaic virus) promoter showed proper expression patterns. We also confirmed Mendelian segregation of the HPT transgene in the Rl generation.n.

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'Hyowon6', a Mid-Late Maturing Rice Variety with High Quality (고품질 중만생 벼 품종 '효원6호')

  • Im, Da-Eun;San, Mar Lar;Jang, Seong-Gyu;Park, So-Yeon;Jin, Sang-Hyeon;Kim, Yong Chul;Ham, Tae-Ho;Kwon, Soon-Wook
    • Korean Journal of Breeding Science
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    • v.50 no.4
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    • pp.529-533
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    • 2018
  • Hyowon6, a mid-late maturing, good eating quality rice variety with multi-resistance, was developed by the rice breeding team of Pusan National University in 2013. This variety was derived from a cross between Hwayeong (as a multi-resistant parent) and Koshihikari (as a good eating quality parent). Selection by the pedigree breeding method was carried out until the $F_8$ generation. A promising line, JS14-12-36-8-5-3-1-1-1, was developed and designated as HY103 in 2013. This variety headed on August 22, which was 11 days later than that of Hwayeong. The culm length and panicle length of Hyowon6 was 85.7 cm and 21.1 cm, respectively. The number of panicles per hill was 14.5 and the number of grains per panicle was 101.7. The ratio of ripened grain was approximately 92% and the 1,000-grain weight was approximately 21.36 g for brown rice, which were similar to those of Hwayeong. Hyowon6 was moderately resistant to lodging and also to neck blast, leaf blight, and stripe virus. The glossiness value of Hyowon6 was 83, which was considerably higher than that of Hwayeong.

Evaluation of the Frequency of the IL-28 Polymorphism (rs8099917) in Patients with Chronic Hepatitis C Using Zip Nucleic Acid Probes, Kerman, Southeast of Iran

  • Iranmanesh, Zahra;Mollaie, Hamid Reza;Arabzadeh, Seyed Alimohammad;Zahedi, Mohammad Javad;Fazlalipour, Mehdi;Ebrahimi, Saeede
    • Asian Pacific Journal of Cancer Prevention
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    • v.16 no.5
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    • pp.1919-1924
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    • 2015
  • Polymorphisms in the region of the interleukin IL-28 gene on chromosome 19 have been related with clearance of hepatitis C virus (HCV), a major human pathogen responsible for chronic hepatitis, cirrhosis and hepatocellular carcinoma. About 3% of the world's population is infected with HCV. The long-term response to therapy is influenced by many host and viral factors, and recent evidence has indicated that some host genetic polymorphisms related to IL-28 are the most powerful predictors of virological response in patients with HCV. This study assessed frequency of the IL-28 polymorphism (rs8099917) in 50 patients (39 men and 11 women) with chronic hepatitis C using ZNA probe real time PCR new method. All patients were tested for genotype of HCV and the HCV viral load. In parallel, the levels of SGOT, SGPT and ALK enzymes were assessed. Treatment using Peg-interferon alpha with ribavirin was conducted for patients and subsequently samples were collected to detect any change in viral load or liver enzyme rates. The overall frequency of the TT allele is 74%, TG allele 20% and GG allele 6% and the percent of patients who had T allele was 84%. Clear reduction in viral load and liver enzymes was reported in patients with the T allele. Especially for genotype 1 which is relatively resistant to treatment, these alleles may have a role in this decline. In conclusion, we showed that IL-28 polymorphism rs8099917 strongly predicts virological response in HCV infection and that real-time PCR with Zip nucleic acid probes is a sensitive, specific and rapid detection method for detection of SNPs which will be essential for monitoring patients undergoing antiviral therapy.

Characteristics and Yield of Recommended Cultivars by Imported Forage Crop Regional Yield Trials in 2002 I. Early Maturing, Good Qualify, and High Yield of Forage Corn Hybrid, “DK 537 (2002년 사료작물 수입적응성 인증품종의 생육기성 및 수동성 1. 조숙 양질 다수성 사료작물 옥수수 교잡종 “DK 537”)

  • Sung, B.R.;Choi, G.J.;Lim, Y.W.;Lim, Y.C.;Park, K.J.
    • Journal of The Korean Society of Grassland and Forage Science
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    • v.22 no.4
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    • pp.247-252
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    • 2002
  • “DK 537”has been selected by forage crop breeding team at the National Livestock Research Institute(NLRI), RDA and has been determined by the Deliberative Council of the National Agricultural Cooperative Federation(NACF) in 2002, as a new recommended hybrid which is early maturing, good quality and hi호 yield of corn for silage. The characteristics of this hybrid are as follows ; 1. The seed coat is yellow and the mean of tasseling date is 5th of July, included in early maturing hybrid which is one day delayed than check cultivar, DK 501. The culm length is 246cm. It is resistant to lodging because of its low height from surface to ear. 2. DK 537 shows resistance to H. maydis and Maize Black-Streaked Dwarf Virus(MBSDV). It also shows strength to corn borer as much as DK 501 does. 3. Fresh yield. dry matter yield. and TDN per a ha are 50 tons, 16.9 tons, and 11.4 tons respectively. which are almost same yielding level of DK 501. Its percent ear to total dry matter is 50.8% at the same time. Through all these tests, we could make sure at DK 537 hybrid will be recommended as good forage crop.

Lack of TNF-α Gene Polymorphism (rs1799724) Association with Sustained Virological Response in Iranian Patients with Chronic HCV Infection

  • Larijani, Mona Sadat;Bahiraei, Narges;Nikbin, Mehri;Mohajel, Nasir;Rad, Leila Naghizadeh;Baghbani, Fahimeh;Mapar, Maryam;Sadat, Seyed Mehdi
    • Asian Pacific Journal of Cancer Prevention
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    • v.17 no.8
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    • pp.3923-3927
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    • 2016
  • Infection with the hepatitis C virus is a major public health concern which can lead to carcinoma and liver failure. It has been shown that single nucleotide polymorphisms can affect the level of gene activity of tumor necrosis factor (TNF) which has an important role, especially in viral infections which can lead to apaptosis of infected hepatocellular cells. We investigated the impact of three possible genotypes for rs1800629 or A/G single nucleotide polymorphism located downstream of $TNF{\alpha}$ gene promoter in groups of control (n=76) and chronic hepatitis C patients (n=89) focusing on the response to treatment among sensitive and resistant groups. Genomic DNA was extracted from $500{\mu}l$ prepheral whole blood and PCR and RFLP were used to amplify the region of interest and genotyping. With statistical analyzes a p-value <0.05 was considered meaningful. There was no significant difference in distribution of possible three genotypes among healthy individuals and patients (P=0.906, OR=1.194, CI=0.063-22.790). However, the frequency of G allele was higher in patients whereas A allele was more common among healthy individuals (p<0.0001). Further studies with more samples seem to be necessary.

Development of a Sequence Characteristic Amplified Region Marker linked to the L4 Locus Conferring Broad Spectrum Resistance to Tobamoviruses in Pepper Plants

  • Kim, Hyun Jung;Han, Jung-Heon;Yoo, Jae Hyoung;Cho, Hwa Jin;Kim, Byung-Dong
    • Molecules and Cells
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    • v.25 no.2
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    • pp.205-210
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    • 2008
  • To develop molecular markers linked to the $L^4$ locus conferring resistance to tobamovirus pathotypes in pepper plants, we performed AFLP with 512 primer combinations for susceptible (S pool) and resistant (R pool) DNA bulks against pathotype 1.2 of pepper mild mottle virus. Each bulk was made by pooling the DNA of five homozygous individuals from a T10 population, which was a near-isogenic $BC_4F_2$ generation for the $L^4$ locus. A total of 19 primer pairs produced scorable bands in the R pool. Further screening with these primer pairs was done on DNA bulks from T102, a $BC_{10}F_2$ derived from T10 by back crossing. Three AFLP markers were finally selected and designated L4-a, L4-b and L4-c. L4-a and L4-c each underwent one recombination event, whereas no recombination for L4-b was seen in 20 individuals of each DNA bulk. Linkage analysis of these markers in 112 $F_2$ T102 individuals showed that they were each within 2.5 cM of the $L^4$ locus. L4-b was successfully converted into a simple 340-bp SCAR marker, designated L4SC340, which mapped 1.8 cM from the $L^4$ locus in T102 and 0.9 cM in another $BC_{10}F_2$ population, T101. We believe that this newly characterized marker will improve selection of tobamovirus resistance in pepper plants by reducing breeding cost and time.

Establishment of Immotalized Human Gingival Fibroblast Cell Lines (불멸화된 치은 섬유아 세포주의 확립)

  • Song, Jae-Bong;Kim, Hyun-A;Hyun, Ha-Na;Kim, Eun-Cheol;You, Hyung-Keun;Shin, Hyung-Shik
    • Journal of Periodontal and Implant Science
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    • v.32 no.3
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    • pp.603-614
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    • 2002
  • Human gingival fibroblasts have proven to useful as a species specific cell culture system in various system on periodontal disease and regeneration. However, their use is limited, since they are hard to obtain and lifespan is short due to replicative senescence. To overcome these disadvantages, we transfected primary human gingival fibroblasts by the E6 and E7 genes of the Human papilloma virus(HPV) 16. The full length of HPV 16 E6 and E7 was cloned from the pBR322 into BamHl and Sal I of a pBabe vector including hygromycin B resistance. Before pBabeE6/E7 plasmid transfection, peak 8 GFP including G418 resistance was transfected into primary GF to check the transfection efficency. PBabe E6/E7 plasmid was transfected using Lipofectamine plus following manufacter's instruction into primary normal human gingival fibroblasts in 60mm dishes with FBS free DMEM. After 2 days of transfection, the cells were treated with hygromycin for 2 weeks until the transfected control cells died. The resulting hygromycin resistant colonies were pooled, and clonned, and sucessful transfection was established for immortalized gingival fibroblast cell lines. Immoralized GF cells showed stellate shape, that is similar to that of orange grains, and more rapid growth and higher proliferation than that of primary gingival fibroblasts. This cell lines overcame crisis and could be cultured over 30 subcultured, could be use for three dimentional culture, epithelial-mesenchymal interaction study.