• Title/Summary/Keyword: Sequence Selection

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Selection of parental monokaryons from Korean Hypsizigus marmoreus by protoplast regeneration (원형질체 재생을 통한 느티만가닥버섯 단핵균주 선발)

  • Oh, Youn-Lee;Kong, Won-Sik;Jang, Kab-Yeul;Shin, Pyung-Gyun;Kim, Eun-Sun;Oh, Min ji;Choi, In-Geol
    • Journal of Mushroom
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    • v.13 no.3
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    • pp.270-273
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    • 2015
  • Hypsizigus marmoreus is commercially the most important edible mushroom in Japan. This mushroom is usually cultivated for a longer period (about 85~120 days) than other mushroom. In order to develop a new cultivar that has a shortened cultivation period, the genome analysis of this strain has been considered. This study aims to obtain parental monokaryotic strains reproducing 'Haemi' cultivar in Hypsizigus marmoreus for reference genome sequencing. The mycelia were cultured in MCM and MYG media for various incubation periods. Homogenized mycelia were treated with commercial cell wall degrading enzymes to maximize protoplasts production yield from Hypsizigus marmoreus. The greatest number of protoplasts was obtained from mycelia cultured in MCM media for 3 days using Novozyme enzyme. The isolated protoplasts were grown in regeneration agar media after two weeks. Regenerated colonies were picked and moved on separated dishes for microscopic observation. Neohaplonts regenerated from dikayotic strains were identified by the absence of clamp connections. We confirmed that one of monokaryotic strains is a parental strain by crossing with an original compatible strain of 'Haemi' cultivar. This parental strain will be used for reference genome sequence analysis.

Studies on Intracellular Functions of the mas3 Gene in the Fission Yeast, Schizosaccharomyces pombe (분열형 효모에서 mas3 유전자의 세포내 기능 연구)

  • Hwang Mi Ra;Cha Jae Young;Shin Sang Min;Park Jong Kun
    • Journal of Life Science
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    • v.15 no.1 s.68
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    • pp.124-131
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    • 2005
  • The regulation of gene expression plays an important rolet in cell cycle controls. In this study, a novel $mas3^+$ (mitosis associated protein) gene, a homolog of human SMARCADl, was isolated and characterized from a fission yeast Schizosaccharomyces pombe. The overall homology between the helicase proteins of the two species is $87\%$. This DEAD/H box-containing molecule has seven highly conserved sequence regions that allow us to place it in the SNF2 family of the helicase superfamily. Knock-out cell of $mas3^+$ gene was constructed using kanMX6 as a selection marker. Survival of mas3 null mutant exposed to UV or MMS was similar to those of wild type cells. $mas3^+$ expression was lowest at $G_2$ and gradually increased. Cytokinesis of mas3 null mutant was abnormal at $26^{\circ}C\;and\;35^{\circ}C$ and a large number of multi-septate cells were produced. These results indicate that the $mas3^+$ is involved in cytokinesis and cell shape control.

Identification of Quantitative Trait Loci Associated with Leaf Length. Width and Length/width Ratio in Two Recombinant Inbred Lines of Soybean (Glycine max L.) (두 집단의 재조합 근친교잡 계통 (RIL) 콩에서 엽장과 엽폭 및 장폭비와 관련된 양적헝질 유전자좌 분석)

  • Kim, Hyeun-Kyeung;Kang, Sung-Taeg
    • Journal of Life Science
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    • v.14 no.5
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    • pp.821-828
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    • 2004
  • The increasing apparent photosynthetic rate per leaf area may improve seed yield in soybean. Leaf area, length and width are related to the photosynthetic capability of the plant. In this study, two populations derived from the cross of Keunolkong, Shinpaldalkong and Iksanl0 were evaluated with simple sequence repeat (SSR) markers to identify length, width and length/width ratio of leaf. Leaf length/width ratio were significantly negative correlation with leaf width in K/S and K/I populations. In the K/S population, two minor QTLs for leaf length (LL) were found on LG Dlb+W and 1. Two QTLs on LG J and L were related to LL in K/I population. Two and three minor QTLs were identified in leaf width with total phenotypic variation of 13% and 18.04 in K/S and K/I populations, respectively. The leaf length/width ratio, two QTLs on LG I and L, and three QTLs on LG Cl, E and L were related to K/S and K/I populations, respectively. Thus it is assumed that the leaf traits are very much dependent on the genotype used and different breeding approach should be considered for the selection of favorite leaf traits in soybean breeding programs.

Curriculum Components Affecting the Professionalism of Dental Hygienists (치과위생사 전문직업성에 영향을 미치는 교육과정 구성요소)

  • Yang, Seung-Kyeong
    • Journal of dental hygiene science
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    • v.11 no.5
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    • pp.455-462
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    • 2011
  • The purpose of this study was to examine the curriculum components affecting the professionalism of dental hygienists and to explore what improvements should be done for each of the influential curriculum components to boost the belief and attitude of dental hygienists. The subjects in this study were 934 dental hygiene students, 76 dental hygiene professors, and 285 dental hygienists, on whom a survey was conducted in person, by mail and by e-mail from April 10 to June 12, 2009. The collected data from 1,295 respondents were analyzed. First, overall dental hygiene professionalism was predicted by intellectual development, vocational education, socio-culturalization, ethics and creativity, which were educational objectives of curriculum, and that didn't have a close relationship to interpersonal understanding and selfactualization. Second, overall dental hygienist professionalism was predicted by learning interest, learning possibility and the significance of subjects, which were selection criteria of teaching content, and that wasn't closely linked to the validity of subjects and social significance. Third, overall dental hygienist professionalism was predicted by continuity and integrity, which were organizing principles of teaching content, and that didn't have a strong relationship to sequence. Fourth, overall dental hygienist professionalism was predicted by the content of school evaluation and school evaluation criteria, which were evaluation elements of curriculum, and that wasn't closely linked to the method of school evaluation, the content of the national exam, the criteria of the national exam and the method of the national exam.

A Set of Allele-specific Markers Linked to L Locus Resistant to Tobamovirus in Capsicum spp. (고추의 Tobamovirus 저항성 L 유전자좌와 연관된 대립유전자 특이적인 마커 세트)

  • Lee, Jun-Dae;Han, Jung-Heon;Yoon, Jae-Bok
    • Horticultural Science & Technology
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    • v.30 no.3
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    • pp.286-293
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    • 2012
  • The resistance to Tobamovirus in Capsicum spp. has been known to be controlled by five different alleles ($L^0$, $L^1$, $L^2$, $L^3$, and $L^4$) of L locus on the telomere of long arm of pepper chromosome 11. To develop a set of molecular markers differentiating all the alleles of L locus, we used five pepper differential hosts including Capsicum annuum Early California Wonder (ECW, $L^0L^0$), C. annuum Tisana ($L^1L^1$), C. annuum Criollo de Morelos 334 (CM334, $L^2L^2$), Capsicum chinense PI 159236 ($L^3L^3$), and Capsicum chacoense PI 260429 ($L^4L^4$). Developing a series of CAPS or SCAR markers specifically linked to the alleles was allowed by the sequence comparison of PCR amplicons of the $L^3$-linked markers (189D23M, A339, and 253A1R) and BAC sequences (FJ597539 and FJ597541) in the pepper differentials. Genotypes deduced by these markers in 48 out of 53 $F_1$ hybrids of commercial pepper varieties were consistent with their phenotypes by bioassay using Tobamovirus pathotypes ($P_0$, $P_1$, and $P_{1,2$). Consequently, these markers can be useful to differentiate L alleles and for breeding Tobamovirus resistance in pepper with marker-assisted selection.

Polymorphisms of melatonin receptor genes and their associations with egg production traits in Shaoxing duck

  • Feng, Peishi;Zhao, Wanqiu;Xie, Qiang;Zeng, Tao;Lu, Lizhi;Yang, Lin
    • Asian-Australasian Journal of Animal Sciences
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    • v.31 no.10
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    • pp.1535-1541
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    • 2018
  • Objective: In birds, three types of melatonin receptors (MTNR1A, MTNR1B, and MTNR1C) have been cloned. Previous researches have showed that three melatonin receptors played an essential role in reproduction and ovarian physiology. However, the association of polymorphisms of the three receptors with duck reproduction traits and egg quality traits is still unknown. In this test, we chose MTNR1A, MTNR1B, and MTNR1C as candidate genes to detect novel sequence polymorphism and analyze their association with egg production traits in Shaoxing duck, and detected their mRNA expression level in ovaries. Methods: In this study, a total of 785 duck blood samples were collected to investigate the association of melatonin receptor genes with egg production traits and egg quality traits using a direct sequencing method. And 6 ducks representing two groups (3 of each) according to the age at first eggs (at 128 days of age or after 150 days of age) were carefully selected for quantitative real-time polymerase chain reaction. Results: Seven novel polymorphisms (MTNR1A: g. 268C>T, MTNR1B: g. 41C>T, and g. 161T>C, MTNR1C: g. 10C>T, g. 24A>G, g. 108C>T, g. 363 T>C) were detected. The single nucleotide polymorphism (SNP) of MTNR1A (g. 268C>T) was significantly linked with the age at first egg (p<0.05). And a statistically significant association (p<0.05) was found between MTNR1C g.108 C>T and egg production traits: total egg numbers at 34 weeks old of age and age at first egg. In addition, the mRNA expression level of MTNR1A in ovary was significantly higher in late-mature group than in early-mature group, while MTNR1C showed a contrary tendency (p<0.05). Conclusion: These results suggest that identified SNPs in MTNR1A and MTNR1C may influence the age at first egg and could be considered as the candidate molecular marker for identify early maturely traits in duck selection and improvement.

Selection of Lactic Acid Bacteria with Antibacterial Activity for Extension of Kimchi Shelf-life (김치의 저장성 향상을 위한 항균활성 우수 유산균 선발)

  • Choi, Hak-Jong;Kim, Yu Jin;Lee, Na Ra;Park, Hae Woong;Jang, Ja Young;Park, Sung-Hee;Kang, Miran;Kim, Hyun Ju;Lee, Jong-Hee;Lee, Jong-Hoon;Pyun, Yu-Ryang;Kim, Tae-Woon
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.43 no.2
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    • pp.328-332
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    • 2014
  • A survey was conducted on the isolation of lactic acid bacteria with antibacterial activity to extend kimchi shelf-life. Antibacterial activity was tested against bacteria associated with acidification of kimchi, including Lactobacillus plantarum, Pediococcus pentosaceus, and Lactobacillus sakei, using agar-well diffusion assay. Two isolates from kimchi were identified as Lactococcus lactis subsp. lactis and Lactobacillus brevis by 16S rRNA gene sequence analysis and API 50 CHL assay, and they showed antibacterial effects against indicator strains. The isolates displayed acid tolerance at pH 3.5, salt tolerance in 5% NaCl, and growth at $4^{\circ}C$. These result imply that the selected strains might be used to extend kimchi shelf-life as a potential starter.

Morphological and Molecular Characterization of Alternaria Isolates from Solanaceous Crops (가지과 작물에서 분리한 Alternaria 속 균의 형태적, 분자생물학적 특징)

  • Yu, Seung-Hun;Cho, Hye-Sun;Kim, Byung-Ryun;Park, Myung-Soo
    • The Korean Journal of Mycology
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    • v.31 no.2
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    • pp.103-113
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    • 2003
  • Twenty five isolates of Alternaria were obtained from various solanaceous crops in Korea. For all isolates, morphological characteristics of the conidia were determined and compared with those of representative isolates of A. solani and A. tomatophila. A selection of the isolates and the representative Alternaria isolates were evaluated for Pathogenicity to potato, tomato, egg plant and red pepper. Molecular characteristics of 17 isolates of Alternaria inculding the representative isolates were determined using sequence analysis of IRS rDNA and histone H3 gene, and URP-PCR analysis. Based on morphological characteristics, the isolates from the solanaceous crops were grouped as identical or very similar to either A. tomatophila (ATO), A. solani (ASO), and unidentified Alternaria sp. (ASP). Isolates of ASO were moderately pathogenic to all the solanaceous crops tested, but ATO isolates were highly pathogenic to tomato and the ASP isolate was pathogenic only to potato. Among the molecular markers used in this study, the URP-PCR analysis was found to be appropriate for taxonomic resolution of these species. Based on the conidial morphology, pathogenicity test and molecular characteristics, A. tomatophila (early blight of tomato) could be distinguished from A. solani (early blight of potato), and the Alternaria sp. (ASP) from potato, which was closely related to ASO in conidial morphology, was considered as a new species.

Knock-in Efficiency Depending on Homologous Arm Structure of the Knock-in Vector in the Bovine Fibroblasts (체세포에 있어서 Knock-in 벡터 상동영역 구조에 따른 Knock-in 효율)

  • Kim, Se Eun;Park, Da Som;Koo, Deog-Bon;Kang, Man-Jong
    • Reproductive and Developmental Biology
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    • v.41 no.1
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    • pp.7-16
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    • 2017
  • The knock-in efficiency in the fibroblast is very important to produce transgenic domestic animal using nuclear transfer. In this research, we constructed three kinds of different knock-in vectors to study the efficiency of knock-in depending on structure of knock-in vector with different size of homologous arm on the ${\beta}-casein$ gene locus in the somatic cells; DT-A_cEndo Knock-in vector, DT-A_tEndo Knock-in vector I, and DT-A_tEndo Knock-in vector II. The knock-in vector consists of 4.8 kb or 1.06 kb of 5' arm region and 1.8 kb or 0.64 kb of 3' arm region, and neomycin resistance gene(neor) as a positive selection marker gene. The cEndo Knock-in vector had 4.8 kb and 1.8 kb homologous arm. The tEndo Knock-in vector I had 1.06 kb and 0.64 kb homologous arm and tEndo Knock-in vector II had 1.06 kb and 1.8 kb homologous arm. To express endostatin gene as transgene, the F2A sequence was fused to the 5' terminal of endostatin gene and inserted into exon 7 of the ${\beta}-casein$ gene. The knock-in vector and TALEN were introduced into the bovine fibroblast by electroporation. The knock-in efficiencies of cEndo, tEndo I, and tEndo II vector were 4.6%, 2.2% and 4.8%, respectively. These results indicated that size of 3' arm in the knock-in vector is important for TALEN-mediated homologous recombination in the fibroblast. In conclusion, our knock-in system may help to create transgenic dairy cattle expressing human endostatin protein via the endogenous expression system of the bovine ${\beta}-casein$ gene in the mammary gland.

Isolation and Selection of Antagonistic Microbes for Biological Control of Zoysiagrass Large Patch Disease (한국잔디 갈색퍼짐병의 생물학적 방제를 위한 길항미생물의 분리 및 선발)

  • Ma, Ki-Yoon;Kwark, Soo Nyeon;Lee, Geung-Joo
    • Horticultural Science & Technology
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    • v.31 no.6
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    • pp.657-665
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    • 2013
  • A large patch disease caused by Rhizoctonia solani AG2-2 (IV) is a serious problem in Korean lawngrass (Zoysia japonica) sites including golf courses and sports fields in Korea. Antagonistic microorganisms against R. solani AG2-2 (IV) were isolated from various forest and crop soil sources in Southern Korea. Among the 61 isolates, I-009, FRIN-001-1, and YPIN-022 strains showing dramatic inhibition of the mycelial growth of R. solani AG2-2 (IV) in the pairing culture were selected as the most potential antagonistic microorganisms for this study. Based on the 16s RNA sequence comparison, I-009 and FRIN-001-1 isolates were identified as Bacillus spp., while YPIN-022 isolate belongs to the genus Pseudomonas. The greater inhibition (clear) zone between two edges of the selected and pathogenic microbes ranged from 11 to 15 mm in three selections, but the others averaged to 7 mm out of 30 mm distance. In another antifungal test using culture filtrate, those three isolates represented a range of 51.7 to 63.5% suppression potential. The selected isolates also inhibited significantly the stem-segment colonization by R. solani AG2-2 (IV) in vivo test by 28.1%, 43.0%, and 23.7% when inoculated with I-009, FRIN-001-1, and YPIN-022, respectively. The highest antagonistic activity for the large patch disease was demonstrated by the isolate FRIN-001-1, which will be useful for developing a bio-pesticide against Rhizoctonia.