• Title/Summary/Keyword: Polymorphic forms

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Absence of DNA Polymorphisms in Myzus persicae (Homoptera: Aphididae) in Relation to their Host Plants (기주식물 종류에 따른 복숭아혹진딧물(Myzus persicae)의 DNA Polymorphism 비교)

  • H. J. Kim;K. S. Boo;K. H. Cho
    • Korean journal of applied entomology
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    • v.35 no.3
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    • pp.209-215
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    • 1996
  • DNA polymorphisms were analyzed for 8 clones of the green peach aphid, Myzus persicae Sulzer, by random amplified polymorphic DNA-polymerase chain reaction (RAPD-PCR). The insect has different host preferences and was even classified into two different species, M. persicae Sulzer and Myzus nicotinae Blackman by their morphological characters, but this point is still in arguement. To identify the differences between two types of the green peach aphid by RAPD-PCR, the template DNA was extracted from 4 clones each of tobacco-feeding and non-tobacco-feeding forms and one hundred primers of 10-nucleotideslong were tested in PCR. The amplified DNAs were analyzed by agarose gel electrophoresis. Eighty-three primers gave amplified DNA fragments with 1 to 22 in number and 500 to 20,000 base pairs in length, but no amplification was observed in the other 17 primers. The average number of fragment per each amplification was about 13. In the case of 82 out of 83 random primers, band patterns of amplified DNA were identical among 8 clones, even though some differences were noticed in the intensity of specific bands. Polymorphism was detected by only one primer within the tobacco-feeding forms, but not between the two host types. The results did not detect any relationship between RAPD polymorphism and their host preference.

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Molecular Analysis of CAG Repeats at Five Different Spinocerebellar Ataxia loci: Correlation and Alternative Explanations for Disease Pathogenesis

  • Alluri, Ravindra Varma;Komandur, Sreelatha;Wagheray, Avinash;Chaudhuri, Jaydip Ray;Sitajayalakshmi, Sitajayalakshmi;Meena, Angmuthu Kanikannan;Jabeen, Afshan;Chawda, Kamalesh;Subhash, Kaul;Krishnaveni, Alladi;Hasan, Qurratulain
    • Molecules and Cells
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    • v.24 no.3
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    • pp.338-342
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    • 2007
  • Spinocerebellar ataxias (SCAs) are caused by expansion of (CAG)n triplet repeats. These repeats occur as polymorphic forms in general population; however, beyond a threshold size they become pathogenic. The sizes and distributions of repeats at the SCA1, SCA2, SCA3, SCA7 and DRPLA loci were assessed by molecular analysis of 124 unrelated ataxia patients and 44 controls, and the association of larger normal (LN) alleles with disease prevalence was evaluated. Triplet repeat expansions in the disease range were detected in 8% (10/124) of the cases, with the majority having expansion at the SCA1 locus. Normal allele ranges in the cohort studied were similar to the Caucasian and North Indian populations but differed from the Korean and Japanese populations at various loci. The percentage of individuals with LN alleles at the SCA1 and SCA2 loci was higher than reported in Indians, Japanese and Caucasians. LN alleles showed a good correlation with the incidence of SCA1, indicating that SCA1 is the most prevalent ataxia in our population. The majority of cases with clinical symptoms of SCA could not be diagnosed by established CAG repeat criteria, suggesting that there may be an alternative basis for disease pathogenesis: (i) Repeats lower than the normal range may also result in abnormal phenotypes (ii) LN alleles at different loci in the same individual may contribute to symptoms (iii) Exogenous factors may play a role in triggering disease symptoms in individuals with LN alleles (iv) Triplet repeats may reach the disease range in the brain but not in the blood.

Evidence of genome duplication revealed by sequence analysis of multi-loci expressed sequence tagesimple sequence repeat bands in Panax ginseng Meyer

  • Kim, Nam-Hoon;Choi, Hong-Il;Kim, Kyung Hee;Jang, Woojong;Yang, Tae-Jin
    • Journal of Ginseng Research
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    • v.38 no.2
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    • pp.130-135
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    • 2014
  • Background: Panax ginseng, the most famous medicinal herb, has a highly duplicated genome structure. However, the genome duplication of P. ginseng has not been characterized at the sequence level. Multiple band patterns have been consistently observed during the development of DNA markers using unique sequences in P. ginseng. Methods: We compared the sequences of multiple bands derived from unique expressed sequence tagsimple sequence repeat (EST-SSR) markers to investigate the sequence level genome duplication. Results: Reamplification and sequencing of the individual bands revealed that, for each marker, two bands around the expected size were genuine amplicons derived from two paralogous loci. In each case, one of the two bands was polymorphic, showing different allelic forms among nine ginseng cultivars, whereas the other band was usually monomorphic. Sequences derived from the two loci showed a high similarity, including the same primer-binding site, but each locus could be distinguished based on SSR number variations and additional single nucleotide polymorphisms (SNPs) or InDels. A locus-specific marker designed from the SNP site between the paralogous loci produced a single band that also showed clear polymorphism among ginseng cultivars. Conclusion: Our data imply that the recent genome duplication has resulted in two highly similar paralogous regions in the ginseng genome. The two paralogous sequences could be differentiated by large SSR number variations and one or two additional SNPs or InDels in every 100 bp of genic region, which can serve as a reliable identifier for each locus.

Chromosome 22 LD Map Comparison between Korean and Other Populations

  • Lee, Jong-Eun;Jang, Hye-Yoon;Kim, Sook;Yoo, Yeon-Kyeong;Hwang, Jung-Joo;Jun, Hyo-Jung;Lee, Kyu-Sang;Son, Ok-Kyung;Yang, Jun-Mo;Ahn, Kwang-Sung;Kim, Eug-Ene;Lee, Hye-Won;Song, Kyu-Young;Kim, Hie-Lim;Lee, Seong-Gene;Yoon, Yong-Sook;Kimm, Ku-Chan;Han, Bok-Ghee;Oh, Berm-Seok;Kim, Chang-Bae;Jin, Hoon;Choi, Kyoung-O.;Kang, Hyo-Jin;Kim, Young-J.
    • Genomics & Informatics
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    • v.6 no.1
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    • pp.18-28
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    • 2008
  • Single nucleotide polymorphisms (SNPs) are the most abundant forms of human genetic variations and resources for mapping complex genetic traits and disease association studies. We have constructed a linkage disequilibrium (LD) map of chromosome 22 in Korean samples and compared it with those of other populations, including Yorubans in Ibadan, Nigeria (YRI), Centre d'Etude du Polymorphisme Humain (CEPH) reference families (CEU), Japanese in Tokyo (JPT) and Han Chinese in Beijing (CHB) in the HapMap database. We genotyped 4681 of 111,448 publicly available SNPs in 90 unrelated Koreans. Among genotyped SNPs, 4167 were polymorphic. Three hundred and five LD blocks were constructed to make up 18.6% (6.4 of 34.5 Mb) of chromosome 22 with 757 tagSNPs and 815 haplotypes (frequency $\geq$ 5.0%). Of 3430 common SNPs genotyped in all five populations, 514 were monomorphic in Koreans. The CHB + JPT samples have more than a 72% overlap with the monomorphic SNPs in Koreans, while the CEU + YRI samples have less than a 38% overlap. The patterns of hot spots and LD blocks were dispersed throughout chromosome 22, with some common blocks among populations, highly concordant between the three Asian samples. Analysis of the distribution of chimpanzee-derived allele frequency (DAF), a measure of genetic differentiation, Fst levels, and allele frequency difference (AFD) among Koreans and the HapMap samples showed a strong correlation between the Asians, while the CEU and YRI samples showed a very weak correlation with Korean samples. Relative distance as a quantitative measurement based upon DAF, Fst, and AFD indicated that all three Asian samples are very proximate, while CEU and YRI are significantly remote from the Asian samples. Comparative genome-wide LD studies provide useful information on the association studies of complex diseases.

Multiple Confirmation and RAPD-genotyping of Enterobacter sakazakii Isolated from Sunsik (선식에서 분리한 Enterobacter sakazakii의 복합동정 및 RAPD를 이용한 genotyping)

  • Choi, Jae-Won;Kim, Yun-Ji;Lee, Jong-Kyung;Kim, Young-Ho;Kwon, Ki-Sung;Hwang, In-Gyun;Oh, Se-Wook
    • Korean Journal of Food Science and Technology
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    • v.40 no.1
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    • pp.101-105
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    • 2008
  • Enterobacter sakazakii is implicated in severe forms of neonatal infections such as meningitis and sepsis. This organism has been isolated from a wide range of foods, including cheese, vegetables, grains, herbs, and spices, but its primary environment is still unknown. Generally, dried infant milk formula has been epidemiologically identified as the source of E. sakazakii. Sunsik (a powdered mixture of roasted grains and other foodstuffs) is widely consumed in Korea as a side dish or energy supplement. Sunsik is consumed without heat treatment; thus, lacking an additional opportunity to inactivate foodborne pathogens. Therefore, its microbiological safety should be guaranteed. In this study, the prevalence of E. sakazakii was monitored in 23 different sunsik component flours, using FDA recommended methods; but E. sakazakii medium (Neogen) and Chromogenic E. sakazakii medium (Oxoid) were used as the selective media. In total, presumptive E. sakazakii strains were isolated from 8 different sunsik powders. Subsequently, an API 20E test was conducted, and 15 strains from 5 different sunsik flours (sea tangle, brown rice, non-glutinous rice, cheonggukjang, dried anchovy) were confirmed as E. sakazakii. Fifteen strains were again confirmed by PCR amplification, using three different primer sets (tDNA sequence, ITS sequence, 16S rRNA sequence), and compared to ATCC strains (12868, 29004, 29544, 51329). They were once again confirmed by their enzyme production profiles using an API ZYM kit. Finally, RAPD (random amplified polymorphic DNA)-genotyping was carried out as a monitoring tool to determine the contamination route of E. sakazakii during processing.

Development of a CAPS Marker Derived from the Pg-Actin Gene Sequences and RAPD Markers in Platycodon grandiflorum (도라지에서의 RAPD 마커 분석과 Actin 유전자 염기서열에서 유래한 CAPS 분자표지 개발)

  • Kim, Munhwi;Jeong, Eunah;Jeong, Jeongsu;Kwon, Soontae;Jeon, Ikjo;Jeong, Jeong Hag;Lee, Je Min;Yeam, Inhwa
    • Korean Journal of Plant Resources
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    • v.28 no.5
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    • pp.648-655
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    • 2015
  • Balloon flower (Platycodon grandiflorum A. DC.) is a perennial plant of mainly Campanulaceae family, which have been widely used as a food ingredient and herbal medicine in East Asia. Although demands on related products and yearly cultivation area for balloon flower are increasing, diverse fundamental technologies and molecular breeding studies are not very well supported in Platycodons. In this study, 30 random amplification of polymorphic DNA (RAPD) primers were test in an attempt to explore genetic diversities. In addition, sequences information of the actin gene, a well conserved gene encoding a globular protein that forms microfilaments, was retrieved and analyzed. Two actin homologs were recovered; 3.4 kb fragment is a Pg-actin and 1.4 kb fragment is a Pg-actin homolog with 28.6% similarity. We have confirmed that the Pg-actin gene is configured into 4 exons and 3 introns. A single nucleotide polymorphism (SNP), G↔A, was detected on the intron 3, which served as a target for the CAPS marker development. The marker Pg-Actin-Int3 was applied to 32 balloon flower accessions. Balloon flower DNA sequence information generated in this study is expected to contribute to the analysis and molecular breeding and genetic diversity analysis of balloon flowers.

Development of SCAR Markers for the Discrimination of Rhei Radix et Rhizoma and Rhei Undulatai Rhizoma based on the RAPD (RAPD 분석을 통한 대황(大黃)과 종대황(種大黃) 감별용 SCAR 유전자 마커 개발)

  • Moon, Byeong-Cheol;Lee, Young-Mi;Chun, Jin-Mi;Lee, A-Young;Yoon, Tae-Sook;Cheon, Myeong-Sook;Choo, Byung-Kil;Kim, Ho-Kyoung
    • The Korea Journal of Herbology
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    • v.24 no.4
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    • pp.115-120
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    • 2009
  • Objectives : Due to the morphological similarity and frequent occurrence of intermediate forms as well as morphological variations of aerial part, the correct identification between Rhei Radix et Rhizoma and Rhei Undulatai Rhizoma is very difficult. To develop a reliable method for correct identification and improving the quality standards of Rhei Radix et Rhizoma and Rhei Undulatai Rhizoma, we analyzed RAPD and developed SCAR marker. Methods : To amplify target DNA at the genomic level, 32 Operon 10-mer random primers were applied with four Rheum species, R. officinale, R. palmatum, R. tanguticum and R. undulatum. The nucleotide sequences were determined and species-specific primers were prepared depending on the species-specific RAPD amplicons after subcloned into the pGEM-Teasy vector. To develop the SCAR markers, species-specific PCR amplification and multiplex-PCR were carried out using the single species-specific primer pairs and combinations of them, respectively. Results : We used RAPD analysis of four Rheum plant species to obtain several species-specific RAPD amplicons. From nucleotide sequences of these RAPD amplicons, we developed two SCAR markers that amplified 314 bp and 390 bp DNA fragments in only R. undulatum but not in R. officinale, R. palmatum, R. tanguticum and R. undulatum, for distinguishing Rhei Undulatai Rhizoma and Rhei Radix et Rhizoma. Furthermore, we established SCAR markers for the simultaneous discrimination of the three species within a single reaction by using multiplex-PCR. Conclusions : These genetic markers can be used for the efficient discrimination of plants species and commercial herbal medicines between Rhei Undulatai Rhizoma and Rhei Radix et Rhizoma, to ultimately prevent indiscriminate distribution and prescription of these herbal medicines.

Enzymatic synthesis of structured lipids containing conjugated linolenic acids extracted from pomegranate seed oil and their physicochemical characteristics (석류 종자유로부터 얻어진 Conjugated Linolenic Acid를 함유한 기능성 고체지의 효소적 합성 및 이화학적 특성 연구)

  • Lee, Koo;Shin, Jung-Ah;Lee, Ki-Teak
    • Korean Journal of Agricultural Science
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    • v.39 no.3
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    • pp.395-405
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    • 2012
  • Lipozyme TL IM-catalyzed esterification was carried out to produce functional hard fat (structured lipid, SL) using palm stearin (PS) and hydrolysate of pomegranate seed oil (HPSO) of 1:6 molar ratio. HPSO contained conjugated linolenic acid (CLnA, about 80%). The reaction was performed at non-solvent system and solvent (n-hexane) system using Lipozyme TL IM (10% of total substrates, w/w) for 12, 24, and 72 hr in a shaking water bath ($55^{\circ}C$ and 185 rpm), respectively. SL synthesized in non-solvent system (NH-SL) and SL synthesized in n-hexane system (H-SL) were refined after deacidification, respectively. Their physicochemical properties were compared to obtain desirable functional hard fat. The content of CLnA in NH-SL increased from 34.38% to 40.63% with increasing reaction time. Similar results also observed in H-SL resulting in 36.81~45.83% of CLnA. In triacylglycerol (TAG) composition, the main molecules of LnLnLn (Ln=linolenic acid, PN=36) and the LnLnP (P=palmitic acid, PN=40) were newly synthesized in NH-SL and H-SL with increasing reaction time. After 72 hr reaction, iodine values of NH-SL (136.49) and H-SL (140.37) showed high values because of the high content of CLnA. Solid fat index (SFI) in NH-SL was higher than that in H-SL at each measured temperature. The predominant polymorphic forms of NH-SL and H-SL obtained after esterification for 72 hr were the desirable crystalline structure of the ${\beta}$' form.