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Studies on the MC1R Gene Frequencies in Landrace, Large White, Duroc and Jeju Native Black Pigs (랜드레이스, 대요크셔, 듀록 및 제주 흑돈의 Melanocortin 1 Receptor(MC1R) 유전자의 유전자형 분석)

  • Cho, I.C.;Lee, J.G.;Jung, J.G.;Yang, B.S.;Kang, S.Y.;Kim, B.W.
    • Journal of Animal Science and Technology
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    • v.44 no.2
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    • pp.207-212
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    • 2002
  • This study was conducted to investigate the genotypes and frequencies of Melanocortin 1 Receptor(MC1R) genes in pigs which plays a central role in regulation of eumelanin (black/brown) and phaeomelanin(red/yellow) pigment synthesis within the mammalian melanocytes. Four different breeds of pigs(20 Landrace, 20 Yorkshire, 20 Duroc, and 93 Jeju native black pigs) were used and PCR-RFLP analysis of MC1R gene was also carried out. Two regions of MC1R genes (428bp and 405bp) were amplified using two specific primers (MERL1-EPIG2, EPIG1-EPIG3), respectively and MC1R allele were determined using 2 restriction enzymes (BspHⅠ, AccⅡ). The results of this experiment indicated that MC1R allelic type in Landrace, Large Yorkshire and Duroc were MC1R *2 (Ep), MC1R *2 (Ep), MC1R *4 (e), respectively. However, various allelic types of MC1R genes were detected in Jeju native black pigs. MC1R allelic type of Jeju black pigs was MC1R*2 type as in Meishan and Large black breeds or MC1R*3 type as in Hampshire and Berkshire breeds and the gene frequencies of ED1 and ED2 were 0.554 and 0.446 in average.

Genetic Relationship between Populations and Analysis of Genetic Structure in the Korean Native Chicken and the Endemic Chicken Breeds (한국재래닭 및 토착화 품종간의 유연 관계 및 유전 특성 분석)

  • Oh, J.D.;Kang, B.S.;Kim, H.K.;Park, M.N.;Chae, E.J.;Seo, O.S.;Lee, H.K.;Jeon, G.J.;Kong, H.S.
    • Korean Journal of Poultry Science
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    • v.35 no.4
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    • pp.361-366
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    • 2009
  • The purpose of this study was to assess the genetic variation and establish the relationship amongst breeds and strains using 7 chicken specific microsatellite markers. A total of 317 DNA samples from four Korean native chicken (KNC) strains (KR: Korean Native Red chicken strain, KY: Korean Native Yellow chicken strain, KL: Korean Native Black chicken strain, KO: Ogol chicken strain) and three introduced endemic chicken breeds (LE: Leghorn chicken breed, RI: Rhode Island Red chicken breed, CO: Cornish chicken breed). The size of microsatellite markers was decided using GeneMapper Software (v.4.0) after being analyzed using an ABI 3130 Genetic Analyzer. Frequencies of microsatellites markers were used to estimate heterozygosities and genetic distances. The lowest distance (0.074) was observed between the KY and KL breeds and the highest distance (0.779) between the KL and LE breeds. The KNC strains (KR, KY, KL) have comparatively near genetic distance each other. On the other side, each individual was not ramified to different groups and were spread evenly in phylogenetic dendrogram about all the KNC of each strain populations. But the endemic breed populations (LE, RI, CO) were ramified to different groups. The microsatellite polymorphism data were shown to be useful for assessing the genetic relationship between Korean native strains and other foreign breeds.

PERIPHERAL NERVE REGENERATION USING POLYGLYCOLIC ACID CONDUIT AND BRAIN-DERIVED NEUROTROPHIC FACTOR GENE TRANSFECTED SCHWANN CELLS IN RAT SCIATIC NERVE (BDNF 유전자 이입 슈반세포와 PGA 도관을 이용한 백서 좌골신경 재생에 관한 연구)

  • Choi, Won-Jae;Ahn, Kang-Min;Gao, En-Feng;Shin, Young-Min;Kim, Yoon-Tae;Hwang, Soon-Jeong;Kim, Nam-Yeol;Kim, Myung-Jin;Jo, Seung-Woo;Kim, Byung-Soo;Kim, Yun-Hee;Kim, Soung-Min;Lee, Jong-Ho
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • v.30 no.6
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    • pp.465-473
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    • 2004
  • Purpose : The essential triad for nerve regeneration is nerve conduit, supporting cell and neurotrophic factor. In order to improve the peripheral nerve regeneration, we used polyglycolic acid(PGA) tube and brain-derived neurotrophic factor(BDNF) gene transfected Schwann cells in sciatic nerve defects of SD rat. Materials and methods : Nerve conduits were made with PGA sheet and outer surface was coated with poly(lactic-co-glycolic acid) for mechanical strength and control the resorption rate. The diameter of conduit was 1.8mm and the length was 17mm Schwann cells were harvested from dorsal root ganglion(DRG) of SD rat aged 1 day. Schwann cells were cultured on the PGA sheet to test the biocompatibility adhesion of Schwann cell. Human BDNF gene was obtained from cDNA library and amplified using PCR. BDNF gene was inserted into E1 deleted region of adenovirus shuttle vector, pAACCMVpARS. BDNF-adenovirus was multiplied in 293 cells and purified. The BDNF-Adenovirus was then infected to the cultured Schwann cells. Left sciatic nerve of SD rat (250g weighing) was exposed and 14mm defects were made. After bridging the defect with PGA conduit, culture medium(MEM), Schwann cells or BDNF-Adenovirus infected Schwann cells were injected into the lumen of conduit, respectively. 12 weeks after operation, gait analysis for sciatic function index, electrophysiology and histomorphometry was performed. Results : Cultured Schwann cells were well adhered to PGA sheet. Sciatic index of BDNF transfected group was $-53.66{\pm}13.43$ which was the best among three groups. The threshold of compound action potential was between 800 to $1000{\mu}A$ in experimental groups which is about 10 times higher than normal sciatic nerve. Conduction velocity and peak voltage of action potential of BDNF group was the highest among experimental groups. The myelin thickness and axonal density of BDNF group was significantly greater than the other groups. Conclusion : BDNF gene transfected Schwann cells could regenerate the sciatic nerve gap(14mm) of rat successfully.

Genetic Variations of Chicken MC1R Gene and Associations with Feather Color of Korean Native Chicken (KNC) 'Woorimatdag' (토종 '우리맛닭' 부계 및 실용계에서 MC1R 유전자 변이 및 모색과의 연관성 분석)

  • Park, Mi Na;Kim, Tae-Hun;Lee, Hyun-Jeong;Choi, Jin Ae;Heo, Kang-Nyeong;Kim, Chong-Dae;Choo, Hyo-Jun;Han, Jae-Yong;Lee, Taeheon;Lee, Jun-Heon;Lee, Kyung-Tai
    • Korean Journal of Poultry Science
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    • v.40 no.2
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    • pp.139-145
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    • 2013
  • There are several loci controlling the feather color of birds, of which one of the most studied is Extended black (E) encoding the melanocortin 1-receptor (MC1R). Mutations in this gene affect the relative distribution of eumelanin, phaeomelanin. The association of feather color and sequence polymorphism in the melanocortin 1-receptor (MC1R) gene was investigated using Korean native chicken H breed (H_PL) and 'Woorimatdag' commercial chickens (Woorimatdag_CC). In order to correlate gene mutation to Korean native chicken feather color, single nucleotide polymorphism (SNP) from MC1R gene sequence were investigated. A total of 307 birds from H_PL and Woorimatdag_CC were used. H_PL have black, black-brown feather color and Woorimatdag_CC have black with brown spots or brown with black spots. There are 6 SNPs in MC1R gene, locus T69C, C212T, A274G, G376A, G636A, T637C. 3 SNPs are nonsynonymous that change amino acid. But it is difficult to find correlation of feather color and polymorphisms. It will be needed to increase the population of Korean native chicken H breed and correlation analysis of genetic variation with feather colors.

Development of pSJE6c, an Expression Vector for Kimchi Lactic Acid Bacteria, and Heterologous Gene Expression Using the Vector (김치유산균용 발현벡터 pSJE6c 개발과 이를 이용한 외래 유전자 발현)

  • Lee, Kang-Wook;Park, Ji-Yeong;Lee, Ji-Yeon;Lee, Hwang-A;Baek, Chang-Un;Jo, Hyeon-Deok;Kim, Joo-Yeon;Kwon, Gun-Hee;Chun, Ji_Yeon;Kim, Jeong-Hwan
    • Microbiology and Biotechnology Letters
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    • v.37 no.4
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    • pp.389-398
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    • 2009
  • Development of expression vectors is important for the basic and applied researches on kimchi LAB (lactic acid bacteria). An expression vector, pSJE6c was constructed by inserting P6C promoter sequence from Lactococcus lactis into pSJE, a shuttle vector for E. coli and Leuconostoc species. To test the efficiency of pSJE6c, aga ($\alpha$-galactosidase) and lacZ ($\beta$-galactosidase) genes were expressed in Lactobacillus brevis 2.14. Compared to the pSJE, expression levels of both genes were increased, indicating P6C promoter was better than indigenous promoters. Enzyme activities of L. brevis cells harboring pSJE6caga (pSJE6c with aga) or pSJE6Z (pSJE6c with lacZ) were 1.5-2 fold higher than those with pSJEaga (pSJE with aga) or pSJEZ (pSJE with lacZ). More RNA transcripts were detected in cells harboring pSJE6c based recombinant plasmid. The results indicated that heterologous gene expressions in kimchi LAB could be improved significantly by use of efficient expression vectors.

Construction and characterization of heterozygous diploid Escherichia coli (2배체 대장균의 제조와 그 특성)

  • Jung, Hyeim;Lim, Dongbin
    • Korean Journal of Microbiology
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    • v.52 no.4
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    • pp.406-414
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    • 2016
  • Among 6 leu codons, CUG is the most frequently used codon in E. coli. It is recognized by leu-tRNA(CAG) encoded by four genes scattered on two chromosomal loci (leuT and leuPQV ). In the process of constructing a strain with no functional leu-tRNA (CAG) gene on chromosome, we made two mutant strains separately, one on leuPQV locus (${\Delta}leuPQV$), and the other on leuT locus [$leuT^*$(GAG)], where the anticodon of leuT was changed from CAG to GAG, thereby altering its recognition codon from CUG to CUC. We attempted to combine these two mutations by transduction using $leuT^*$(GAG) strain as a donor and ${\Delta}leuPQV$ strain as a recipient. Large and small colonies appeared from this transduction. From PCR and DNA sequencing, large colony was confirmed to be the reciprocal recombinant as expected, but the small colonies contained both mutant $leuT^*$(GAG) and wild type leuT (CAG) genes in the cell. This heterozygous diploid strain did not show any unusual morphology under microscopic observation, but, interestingly, it showed a linear growth curve in rich medium with much slower growth rate than wild type cell. It always formed homogenous small colonies in the selection medium, but, when there was no selection, it readily segregated into $leuT^*$(GAG) and leuT (CAG). From these observations, we suggested that the strain with both $leuT^*$(GAG) and leuT (CAG) genes was not a partial diploid (merodiploid), but a full diploid cell having two different chromosomes. We proposed a model explaining how such a heterozygous diploid cell was formed and how and why its growth showed a linear growth curve.

Immune responses of hepatitis B vaccination among very low birth weight infant (극소 저출생체중아의 영아기 B형 간염 항체 생성률 조사)

  • Kim, Young-Deuk;Han, Myung-Ki;Kim, Ai-Rhan E.;Kim, Ki-Soo;Pi, Soo-Young
    • Clinical and Experimental Pediatrics
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    • v.49 no.8
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    • pp.857-863
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    • 2006
  • Purpose : To evaluate the immunogenicity of hepatitis B vaccine among very low birth weight infants(VLBWI) who were vaccinated at 0, 1, 6 months of chronological age and to determine the factors associated with antibody formations. Methods : A total of 243 VLBWI admitted to Seoul and Gangneung Asan Medical Center neonatal intensive care units from 1997 to 2004 were included. Of 243, 13 infants were born to HBs Ag positive mother. All infants were given DNA recombinant vaccine at 0, 1, and 6 months of chronological age. Infants born to HBs Ag positive mothers received hepatitis B immunoglobulin at birth and a total of 4 doses of vaccinations. An antibody level over 10 mIU/mL, tested at 3-4 months after last vaccination, was regarded as a positive seroconversion. Results : The seroconversion rates were 84.4 percent and 84.5 percent for VLBWI and extremely low birth weight infants(ELBWI), respectively. Of 28 seronegative infants who were given revaccinations, 60.7 percent seroconverted, resulting in 95.3 percent, 97.5 percent seroconversion rates for VLBWI and ELBWI, respectively. 76.9 percent of infants born to HBsAg positive mothers seroconverted and none became hepatitis B carriers. Factors such as gestational age, sex, various neonatal illness, and kinds of vaccinations did not influence the formation of the hepatits B antibody, however, the higher the weight at time of first vacciation yielded better seroconversion rate. Conclusion : Revaccination of seronegative VLBWI after 3 doses of hepatitis B vaccinaton is very effective. Therefore, testing the immune status after the hepatitis B vaccination, a practice not routinely done, is highly recommended.

Review of Anti-Leukemia Effects from Medicinal Plants (항 백혈병작용에 관련된 천연물의 자료조사)

  • Pae Hyun Ock;Lim Chang Kyung;Jang Seon Il;Han Dong Min;An Won Gun;Yoon Yoo Sik;Chon Byung Hun;Kim Won Sin;Yun Young Gab
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.17 no.3
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    • pp.605-610
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    • 2003
  • According to the Leukemia and Lymphoma Society, leukemia is a malignant disease (cancer) that originates in a cell in the marrow. It is characterized by the uncontrolled growth of developing marrow cells. There are two major classifications of leukemia: myelogenous or lymphocytic, which can each be acute or chronic. The terms myelogenous or lymphocytic denote the cell type involved. Thus, four major types of leukemia are: acute or chronic myelogenous leukemia and acute or chronic lymphocytic leukemia. Leukemia, lymphoma and myeloma are considered to be related cancers because they involve the uncontrolled growth of cells with similar functions and origins. The diseases result from an acquired (not inherited) genetic injury to the DNA of a single cell, which becomes abnormal (malignant) and multiplies continuously. In the United States, about 2,000 children and 27,000 adults are diagnosed each year with leukemia. Treatment for cancer may include one or more of the following: chemotherapy, radiation therapy, biological therapy, surgery and bone marrow transplantation. The most effective treatment for leukemia is chemotherapy, which may involve one or a combination of anticancer drugs that destroy cancer cells. Specific types of leukemia are sometimes treated with radiation therapy or biological therapy. Common side effects of most chemotherapy drugs include hair loss, nausea and vomiting, decreased blood counts and infections. Each type of leukemia is sensitive to different combinations of chemotherapy. Medications and length of treatment vary from person to person. Treatment time is usually from one to two years. During this time, your care is managed on an outpatient basis at M. D. Anderson Cancer Center or through your local doctor. Once your protocol is determined, you will receive more specific information about the drug(s) that Will be used to treat your leukemia. There are many factors that will determine the course of treatment, including age, general health, the specific type of leukemia, and also whether there has been previous treatment. there is considerable interest among basic and clinical researchers in novel drugs with activity against leukemia. the vast history of experience of traditional oriental medicine with medicinal plants may facilitate the identification of novel anti leukemic compounds. In the present investigation, we studied 31 kinds of anti leukemic medicinal plants, which its pharmacological action was already reported through many experimental articles and oriental medical book: 『pharmacological action and application of anticancer traditional chinese medicine』 In summary: Used leukemia cellline are HL60, HL-60, Jurkat, Molt-4 of human, and P388, L-1210, L615, L-210, EL-4 of mouse. 31 kinds of anti leukemic medicinal plants are Panax ginseng C.A Mey; Polygonum cuspidatum Sieb. et Zucc; Daphne genkwa Sieb. et Zucc; Aloe ferox Mill; Phorboc diester; Tripterygium wilfordii Hook .f.; Lycoris radiata (L Her)Herb; Atractylodes macrocephala Koidz; Lilium brownii F.E. Brown Var; Paeonia suffruticosa Andr.; Angelica sinensis (Oliv.) Diels; Asparagus cochinensis (Lour. )Merr; Isatis tinctoria L.; Leonurus heterophyllus Sweet; Phytolacca acinosa Roxb.; Trichosanthes kirilowii Maxim; Dioscorea opposita Thumb; Schisandra chinensis (Rurcz. )Baill.; Auium Sativum L; Isatis tinctoria, L; Ligustisum Chvanxiong Hort; Glycyrrhiza uralensis Fisch; Euphorbia Kansui Liou; Polygala tenuifolia Willd; Evodia rutaecarpa (Juss.) Benth; Chelidonium majus L; Rumax madaeo Mak; Sophora Subprostmousea Chunet T.ehen; Strychnos mux-vomical; Acanthopanax senticosus (Rupr.et Maxim.)Harms; Rubia cordifolia L. Anti leukemic compounds, which were isolated from medicinal plants are ginsenoside Ro, ginsenoside Rh2, Emodin, Yuanhuacine, Aleemodin, phorbocdiester, Triptolide, Homolycorine, Atractylol, Colchicnamile, Paeonol, Aspargus polysaccharide A.B.C.D, Indirubin, Leonunrine, Acinosohic acid, Trichosanthin, Ge 132, Schizandrin, allicin, Indirubin, cmdiumlactone chuanxiongol, 18A glycyrrhetic acid, Kansuiphorin A 13 oxyingenol Kansuiphorin B. These investigation suggest that it may be very useful for developing more effective anti leukemic new dregs from medicinal plants.

Effect of Soybean Meal and Soluble Starch on Biogenic Amine Production and Microbial Diversity Using In vitro Rumen Fermentation

  • Jeong, Chang-Dae;Mamuad, Lovelia L.;Kim, Seon-Ho;Choi, Yeon Jae;Soriano, Alvin P.;Cho, Kwang Keun;Jeon, Che-Ok;Lee, Sung Sil;Lee, Sang-Suk
    • Asian-Australasian Journal of Animal Sciences
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    • v.28 no.1
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    • pp.50-57
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    • 2015
  • This study was conducted to investigate the effect of soybean meal (SM) and soluble starch (SS) on biogenic amine production and microbial diversity using in vitro ruminal fermentation. Treatments comprised of incubation of 2 g of mixture (expressed as 10 parts) containing different ratios of SM to SS as: 0:0, 10:0, 7:3, 5:5, 3:7, or 0:10. In vitro ruminal fermentation parameters were determined at 0, 12, 24, and 48 h of incubation while the biogenic amine and microbial diversity were determined at 48 h of incubation. Treatment with highest proportion of SM had higher (p<0.05) gas production than those with higher proportions of SS. Samples with higher proportion of SS resulted in lower pH than those with higher proportion of SM after 48 h of incubation. The largest change in $NH_3$-N concentration from 0 to 48 h was observed on all SM while the smallest was observed on exclusive SS. Similarly, exclusive SS had the lowest $NH_3$-N concentration among all groups after 24 h of incubation. Increasing methane ($CH_4$) concentrations were observed with time, and $CH_4$ concentrations were higher (p<0.05) with greater proportions of SM than SS. Balanced proportion of SM and SS had the highest (p<0.05) total volatile fatty acid (TVFA) while propionate was found highest in higher proportion of SS. Moreover, biogenic amine (BA) was higher (p<0.05) in samples containing greater proportions of SM. Histamines, amine index and total amines were highest in exclusive SM followed in sequence mixtures with increasing proportion of SS (and lowered proportion of SM) at 48 h of incubation. Nine dominant bands were identified by denaturing gradient gel electrophoresis (DGGE) and their identity ranged from 87% to 100% which were mostly isolated from rumen and feces. Bands R2 (uncultured bacterium clone RB-5E1) and R4 (uncultured rumen bacterium clone L7A_C10) bands were found in samples with higher proportions of SM while R3 (uncultured Firmicutes bacterium clone NI_52), R7 (Selenomonas sp. MCB2), R8 (Selenomonas ruminantium gene) and R9 (Selenomonas ruminantium strain LongY6) were found in samples with higher proportions of SS. Different feed ratios affect rumen fermentation in terms of pH, $NH_3$-N, $CH_4$, BA, volatile fatty acid and other metabolite concentrations and microbial diversity. Balanced protein and carbohydrate ratios are needed for rumen fermentation.

Etiological Properties and Coat Protein Gen Analysis of Potato Virus Y Occuring in Potatoes of Korea (우리나라 감자에 발생하는 PVY의 병원학적 특성 및 외피단백질 유전자 분석)

  • ;Richard M. Bostock
    • Proceedings of the Korean Society of Plant Pathology Conference
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    • 1995.06b
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    • pp.77-96
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    • 1995
  • To obtain basic informations for the improvement of seed potato production in Korea, some etiological properties of potato virus Y(PVY) distributed in the major seed potato production area(Daekwanryeong) were characterized, and the nucleotide and amino acid sequences of the coat protein gene of the PVY strains isolated were analyzed. PVY strains in Daekwonryeong, an alpine area, were identified to be two strains, PVYo and PVYN by symptoms of indicator plants, and their distribution in potato fields was similar. Major symptom on potato varieties by PVY was grouped as either mosaic alone or mosaic accompanied with veinal necrosis in the lower leaves. The symptom occurrence of the two symptoms was similar with Irish Cobbler, but Superior showed a higher rate of mosaic symptom than the other. The PVY strain which was isolated from potato cv. Superior showing typical mosaic symptoms produced symptoms of PVY-O on the indicator plants of Chenopodium amaranticolor, Nicotiana tabacum cv. Xanthi nc and Physalis floridana, but no symptom o Capsicum annum cv. Ace. Moreover, results from the enzyme-linked immunosorbent assay with monoclonal and polyclonal antibodies showed that the isolated PVY reacts strongly with PYV-O antibodies but does not react specifically with PVY-T antibodies. The purified virus particles were flexious with a size of 730$\times$11nm. On the basis of the above characteristics, the strain was identified to be a PVY-O and named as of PVY-K strain. The flight of vector aphids was observed in late May, however, the first occurrence of infected plants was in mid June with the bait plants surrounded with PVY-infected potato plants and early July with the bait plants surrounded with PVY-free potato plants. PVY infection rates by counting symptoms on bait plants (White Burley) were 1.1% with the field surrounded with PVY-free potato plants and 13.7% the fields surrounded with PVY-infected potato plants, showing the effect of infection pressure. The propagated PVY-K strain on tobacco(N. sylvestris) was purified, and the RNA of the virus was extracted by the method of phenol extraction. The size of PVY-K RNA was measured to be 9, 500 nucleotides on agarose gel electrophoresis. The double-stranded cDNAs of PVY-K coat protein(CP) gene derived by the method of polymerase chain reaction were transformed into the competent cells of E. coli JM 109, and 2 clones(pYK6 and pYK17) among 11 clones were confirmed to contain the full-length cDNA. Purified plasmids from pYK17 were cut with Sph I and Xba I were deleted with exonuclease III and were used for sequencing analysis. The PVY-K CP gene was comprised of 801 nucleotides when counted from the clevage site of CAG(Gln)-GCA(Ala) to the stop codon of TGA and encoded 267 amino acids. The molecular weight of the encoded polypeptides was calculated to be 34, 630 daltons. The base composition of the CP gene was 33.3% of adenine, 25.2% of guanine, 20.1% of cytosine and 21.4% of uracil. The polypeptide encoded by PVY-K CP gene was comprised of 22 alanines, 20 threonines, 19 glutamic acids and 18 glycines in order. The homology of nucleotide sequence of PVY-K CP gene with those of PVY-O(Japan), PVY-T(Japan), PVY-TH(Japan), PVYN(the Netherlands), and PVYN(France) was represented as 97.3%, 88.9%, 89.3%, 89.6% and 98.5%, respectively. The amino acid sequence homology of the polypeptide encoded by PVY-K CP gene with those encoded by viruses was represented as 97.4%, 92.5%, 92.9%, 92.9%, and 98.5%, respectively.

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