• Title/Summary/Keyword: gene pool

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Mass production and application of activation tagged hairy root lines for functional genomic of secondary metabolism in ginseng

  • Choi, Dong-Woog;Chung, Hwa-Jee;Ko, Suk-Min;In, Dong-Soo;Song, Ji-Sook;Woo, Sung-Sick;Liu, Jang R.
    • Journal of Plant Biotechnology
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    • v.36 no.3
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    • pp.294-300
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    • 2009
  • Activation tagging that uses T-DNA vectors containing multimerized transcriptional enhancers from the cauliflower mosaic virus (CaMV) 35S gene is a powerful tool to determine gene function in plants. This approach has been successfully applied in screening various types of mutations and cloning the corresponding genes. We generated an activation tagged hairy root pool of ginseng (Panax ginseng C.A. Meyer) in an attempt to isolate genes involved in the biosynthetic pathway of ginsenoside (triterpene saponin), which is known as the major active ingredient of the root. Quantitative and qualitative variation of ginsenoside in activation tagged hairy root lines were profiled using LC/MS. Metabolic profiling data enabled selection of a specific hairy root line which accumulated ginsenoside at a higher level than other lines. The relative expression level of several genes of triterpene biosynthetic pathway in the selected hairy root line was determined by real time RT-PCR. Overall results suggest that the activation tagged ginseng hairy root system described in this study would be useful in isolating genes involved in a complex metabolic pathway from genetically intractable plant species by metabolic profiling.

Genome-wide Association Analyses for Resistance to Phytophthora sojae and Pseudomonas amygdali pv. tabaci in Soybean

  • Hee Jin You;Ruihua Zhao;EunJee Kang;Younghyeon Kim;In Jeong Kang;Sungwoo Lee
    • Proceedings of the Korean Society of Crop Science Conference
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    • 2022.10a
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    • pp.186-186
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    • 2022
  • Phytophthora root and stem rot (PRSR) and wildfire disease (WFD) of soybean are frequently observed in the field of South Korea. The most environmentally friendly way to control PRSR and WFD is to use soybean varieties with resistance to Phytophthora sojae (P. sojae) and Pseudomonas amygdali pv. tabaci. Plant germplasm is an important gene pool for soybean breeding and improvement. In this study, hundreds of soybean accessions were evaluated for the two pathogens, and genome-wide association analyses were conducted using 104,955 SNPs to identify resistance loci for the two pathogens. Of 193 accessions, 46 genotypes showed resistance reaction, while 143 did susceptibility for PRSP. Twenty SNPs were significantly associated with resistance to P. sojae on chromosomes (Chr.) 3 and 4. Significant SNPs on Chr.3 were located within the known Rps gene region. A region on Chr. 4 is considered as a new candidate resistance loci. For evalation of resistance to WFD, 18, 31,74,36 and 34 genotypes were counted by a scale of 1-5, respectively. Five SNP markers on Chrs 9,11,12,17 and 18 were significantly associated with resistance to P. amygdali pv. tabaci. The identified SNPs and genomic regions will provide a useful information for further researches and breeding for resistance to P. sojae and P. amygdali pv. tabaci.

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Isolation and Identification of Acanthamoeba in a Contact Lens Storage Case

  • Moon, Eun-Kyung
    • Biomedical Science Letters
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    • v.18 no.3
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    • pp.324-327
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    • 2012
  • Acanthamoeba is a free-living amoeba that causes human infections, and recently the incidence of amoebic keratitis has increased among contact lens wearers. In order to investigate Acanthamoeba contamination of contact lens storage cases, a short survey was performed on 57 contact lens wearers, and Acanthamoeba was found in one contact lens storage case. To diagnose Acanthamoeba, the 18s small subunit ribosomal DNA (18s rDNA) gene was amplified by polymerase chain reaction (PCR), and subsequently, the isolate was identified as A. lugdunensis. This species was originally isolated from a freshwater pool in France, and was reported recently to be a cause of amoebic keratitis. This observation indicates the need for a large survey to investigate the extent of Acanthamoeba contamination, and suggests that contact lens wearers be aware of the importance of hygiene and of the implications of Acanthamoeba infection.

Isolation of High Quality RNA from Seeds of the Mungbean (Vigna radiata) (녹두 종자의 RNA 분리 방법)

  • Kim, Kyeong-Im;Ku, Ja-Hwan
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.51 no.spc1
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    • pp.274-276
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    • 2006
  • Mungbean (Vigna radiata) is a legume to East Asia that has great potential for development as traditional food and industrial crop. It produces both protein and starch rich grain. The low variability of the existing gene pool of the mungbean limits the use of conventional plant breeding to address this problem. For this purpose, an efficient means of RNA isolation from mungbean seed tissues was developed. The quality of RNA obtained by this method was sufficient for use in RT-PCR and RNA analysis.

Enhancement of Clavulanic Acid Production by Expressing Regulatory Genes in gap Gene Deletion Mutant of Streptomyces clavuligerus NRRL3585

  • Jnawali, Hum Nath;Lee, Hei-Chan;Sohng, Jae-Kyung
    • Journal of Microbiology and Biotechnology
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    • v.20 no.1
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    • pp.146-152
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    • 2010
  • Streptomyces clavuligerus NRRL3585 produces a clinically important $\beta$-lactamase inhibitor, clavulanic acid (CA). In order to increase the production of CA, the glyceraldehyde-3-phosphate dehydrogenase (GAPDH) gene (gap) was deleted in S. clavuligerus NRRL3585 to overcome the limited glyceraldehyde-3-phosphate pool; the replicative and integrative expressions of ccaR (specific regulator of the CA biosynthetic operon) and claR (Lys-type transcriptional activator) genes were transformed together into a deletion mutant to improve clavulanic acid production. We constructed two recombinant plasmids to enhance the production of CA in the gap1 deletion mutant of S. clavuligerus NRRL3585: pHN11 was constructed for overexpression of ccaR-claR, whereas pHN12 was constructed for their chromosomal integration. Both pHN11 and pHN12 transformants enhanced the production of CA by 2.59-fold and 5.85-fold, respectively, compared with the gap1 deletion mutant. For further enhancement of CA, we fed the pHN11 and pHN12 transformants ornithine and glycerol. Compared with the gap1 deletion mutant, ornithine increased CA production by 3.24- and 6.51-fold in the pHN11 and pHN12 transformants, respectively, glycerol increased CA by 2.96- and 6.21-fold, respectively, and ornithine and glycerol together increased CA by 3.72- and 7.02-fold, respectively.

Genetic and morphometric characteristics of Korean wild mice (KWM/Hym) captured at Chuncheon, South Korea

  • Nam, Hajin;Kim, Yoo Yeon;Kim, Boyoung;Yoon, Won Kee;Kim, Hyoung-Chin;Suh, Jun Gyo
    • Laboraroty Animal Research
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    • v.34 no.4
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    • pp.311-316
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    • 2018
  • Laboratory inbred mice are used widely and commonly in biomedical research, but inbred mice do not have a big enough gene pool for the research. In this study, genetic and morphometric analyses were performed to obtain data on the characteristics of a newly developing inbred strain (KWM/Hym) captured from Chuncheon, Korea. All of five Korean wild male mice have the zinc-finger Y (ZfY) gene. Also, all of 19 Korean wild mice used in this analysis have the AKV-type murine leukemia virus gene, indicating that Korean wild mice might be Mus musculus musculus. To identify the genetic polymorphism in KWM/Hym, SNP analysis was performed. In a comparison with 28 SNP markers, there was a considerable difference between KWM/Hym and several inbred strains. The homogeneity between KWM/Hym and the inbred strains was as follows: C57BL/6J (39.3%), BALB/c AJic (42.9%), and DBA/2J (50%). KWM/Hym is most similar to the PWK/PhJ inbred strain (96.4%) derived from wild mice (Czech Republic). To identify the morphometric characteristics of KWM/Hym, the external morphology was measured. The tail ratio of male and female was $79.60{\pm}3.09$ and $73.55{\pm}6.14%$, respectively. KWM/Hym has short and agouticolored hairs and its belly is white with golden hair. Taking these results together, KWM/Hym, a newly developing inbred mouse originated from wild mouse, might be use as new genetic resources to overcome the limitations of the current laboratory mice.

Molecular Mechanisms of 5-Azacytidine-Induced Trifluorothymidine-Resistance In Chinese Hamster V79 Cells

  • Jin Kyong-Suk;Lee Yong-Woo
    • Biomedical Science Letters
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    • v.11 no.2
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    • pp.165-173
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    • 2005
  • A potent demethylating agent, 5-Azacytidine (5-AzaC) has been widely used as in many studies on DNA methylation, regulation of gene expression, and cancer biology. The mechanisms of the demethylating activity were known to be formation of complex between DNA and DNA methyltransferase (MTase), which depletes cellular MTase activity. However, 5-AzaC can also induce hypermethylation of a transgene in a transgenic cell line, G12 cells and it was explained as a result of defense mechanisms to inactivate foreign gene(s) somehow. This finding evoked the question that whether the phenomenon of hypermethylation induced by 5-AzaC is limited to the transgene or it can be occurred in endogenous gene(s). In order to answer the question, mutagenicity test of 5-AzaC and molecular characterization of mutants obtained from the test were performed using an endogenous gene, thymidine kinase (tk) in Chinese hamster V79 cells. When V79 and V79-J3 subclone cells were treated with 1, 2.5 ,5, $10{\mu}M$ of 5-AzaC for 48 hours, their maximum mutant frequencies were revealed as $6\times10^{-3}\;at\;5{\mu}M$(350-fold induction over background) and $8\times10^{-3}\;at\;2.5{\mu}M$ (l,800-fold induction over background) respectively. Since the induction rates were too high to be induced by true mutations, many trifluorothymidine (TFT)-resistant $(TFT^R)$ cells were subjected to Northern blot analysis to check the presence of tk transcripts. Surprisingly, all clones tested possessed the transcripts in a similar level, that implicates the $TFT^R$ phenotype induced by 5-AzaC has not given rise to hypermethylation of the gene in spite of unusually high mutation frequency. In addition, it has shown that the TK activity in the pool of 5-AzaC-induced $TFT^R$ cells has about a half of that in spontaneously-induced $TFT^R$ cells or in non-selected parental V79-J3 cells. This result suggests that the mechanism(s) underlying the TFT-resistance between spontaneously occurred and 5-AzaC-induced cells may be different. These findings have shown that the $TFT^R$ phenotype induced by 5-AzaC has not given rise to hypermethylation of the tk gene, and 5-AzaC may be induced by one or combined pathways among many drug resistance mechanisms. The exact mechanisms for the 5-AzaC-induced $TFT^R$ phenotype remain to elucidate.

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Diversity based Ensemble Genetic Programming for Improving Classification Performance (분류 성능 향상을 위한 다양성 기반 앙상블 유전자 프로그래밍)

  • Hong Jin-Hyuk;Cho Sung-Bae
    • Journal of KIISE:Software and Applications
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    • v.32 no.12
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    • pp.1229-1237
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    • 2005
  • Combining multiple classifiers has been actively exploited to improve classification performance. It is required to construct a pool of accurate and diverse base classifier for obtaining a good ensemble classifier. Conventionally ensemble learning techniques such as bagging and boosting have been used and the diversify of base classifiers for the training set has been estimated, but there are some limitations in classifying gene expression profiles since only a few training samples are available. This paper proposes an ensemble technique that analyzes the diversity of classification rules obtained by genetic programming. Genetic programming generates interpretable rules, and a sample is classified by combining the most diverse set of rules. We have applied the proposed method to cancer classification with gene expression profiles. Experiments on lymphoma cancer dataset, prostate cancer dataset and ovarian cancer dataset have illustrated the usefulness of the proposed method. h higher classification accuracy has been obtained with the proposed method than without considering diversity. It has been also confirmed that the diversity increases classification performance.

Assessment of genetic diversity of Prangos fedtschenkoi (Apiaceae) and its conservation status based on ISSR markers

  • Mustafina, Feruza U.;Kim, Eun Hye;Son, Sung-Won;Turginov, Orzimat T.;Chang, Kae Sun;Choi, Kyung
    • Korean Journal of Plant Taxonomy
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    • v.47 no.1
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    • pp.11-22
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    • 2017
  • Prangos fedtschenkoi (Regel et Schmalh.) Korovin (Apiaceae) is an endemic species for mountainous Middle Asia, which is both a rare and useful plant. Organic extractions from this species are being used in pharmaceutics and cosmetology. In recent years, P. fedtschenkoi distribution area has considerably decreased, presumably, resulting from human activities such as agriculture, construction works, overgrazing and collection from wild for pharmaceutic purposes. Six populations were found in Uzbekistan and their genetic divergence and differentiation were studied with 10 inter-simple sequence repeat (ISSR) markers, selected out of 101. Totally 166 amplified ISSR fragments (loci) were revealed, of which 164 were polymorphic. Relatively moderate level of polymorphism was found at population level with polymorphic bands ranging from 27.71% to 47.59%. Mean P = 39.05%, $N_a=1.40$, $N_e=1.25$, S.I. = 0.21, and $H_e=0.14$ were revealed for all loci across six populations. AMOVA showed higher variation among populations (62%) than within them (38%). The Bayesian model determined 5 clusters, or genetic groups. The posteriori distribution of the Theta II estimator detected full model identifying high inbreeding, intensified by low gene flow (Nm = 0.3954). Mantel test confined population 6 as distinct cluster corresponding to geographic remoteness (R = 0.5137, $p{\leq}0.005$). Results were used as the bases for developing conserve measures to restore populations.

A Simple Stereo Matching Algorithm using PBIL and its Alternative (PBIL을 이용한 소형 스테레오 정합 및 대안 알고리즘)

  • Han Kyu-Phil
    • The KIPS Transactions:PartB
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    • v.12B no.4 s.100
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    • pp.429-436
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    • 2005
  • A simple stereo matching algorithm using population-based incremental learning(PBIL) is proposed in this paper to decrease the general problem of genetic algorithms, such as memory consumption and inefficiency of search. PBIL is a variation of genetic algorithms using stochastic search and competitive teaming based on a probability vector. The structure of PBIL is simpler than that of other genetic algorithm families, such as serial and parallel ones, due to the use of a probability vector. The PBIL strategy is simplified and adapted for stereo matching circumstances. Thus, gene pool, chromosome crossover, and gene mutation we removed, while the evolution rule, that fitter chromosomes should have higher survival probabilities, is preserved. As a result, memory space is decreased, matching rules are simplified and computation cost is reduced. In addition, a scheme controlling the distance of neighbors for disparity smoothness is inserted to obtain a wide-area consistency of disparities, like a result of coarse-to-fine matchers. Because of this scheme, the proposed algorithm can produce a stable disparity map with a small fixed-size window. Finally, an alterative version of the proposed algorithm without using probability vector is also presented for simpler set-ups.