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An Introduction to Microsatellite Development and Analysis (Microsatellite 개발 및 분석법에 대한 소개)

  • Yun Young-Eun;Yu Jeong-Nam;Lee Byoung-Yoon;Kwak Myounghai
    • Korean Journal of Plant Taxonomy
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    • v.41 no.4
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    • pp.299-314
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    • 2011
  • The choice of molecular markers is the first step when selecting experimental plans in the field of population genetics. The popular molecular markers in population genetic studies are mainly allozyme, RAPD, RFLP, AFLP, microsatellite, SNP and ISSR. Among these, microsatellites are frequently found in nuclear, chloroplast and mitochondrial genome, showing a high level of polymorphism and nuclear microsatellites are codominant. Thus, it is a favorable molecular marker for population structure analyses and genetic diversity studies. Microsatellites are composed of tandem repeated 1~6 base pair nucleotide motifs and can be easily amplified by PCR reactions using locus specific primers. Because microsatellites have low cross-species transferability, however, they are only applicable between phylogenetically close species. In wild plants, the lack of genomic information and the high development cost of the microsatellite obstruct the wider use of microsatellites in plant population genetics research. In this review, we introduce the basis for microsatellite markers, the development process, and analytical methods as well as evolutionary models and their applications. In addition, possible genotyping errors which lead to erroneous conclusions are discussed.

Association Study Between Dopamine Transporter Gene 40 bp VNTR and Antipsychotics-Induced Restless Legs Syndrome (도파민 수송체 유전자 다형성과 항정신병약물로 유발된 하지불안증후군의 연관성 연구)

  • Kang, Seung-Gul;Lee, Heon-Jeong;Choi, Jung-Eun;Kim, Leen;Jung, In-Kwa
    • Sleep Medicine and Psychophysiology
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    • v.15 no.1
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    • pp.39-43
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    • 2008
  • Objectives: The pathophysiology of restless legs syndrome (RLS) is not obvious, but many promising theories involve dopaminergic deficiency and genetic causes. The RLS is presumed to occur more frequently among schizophrenic patients who take antipsychotics, most of which blocks the dopamine receptors. This study aimed to investigate whether dopamine transporter gene (DAT1) 40 base pair (bp) variable number of tandem repeat (VNTR) polymorphism is associated with the antipsychotic-induced RLS in schizophrenia. Methods: We determined the diagnosis of RLS among the 190 Korean schizophrenic patients by the diagnostic criteria of the International Restless Legs Syndrome Study Group (IRLSSG). Genotyping was performed for the 40bp VNTR in DAT1 gene using polymerase chain reaction. Results: We separated the schizophrenic patients into 44 patients with RLS and 146 patients without RLS. The genotype and allele frequencies did not differ significantly between two groups. Conclusions: These results suggest that DAT1 gene 40bp VNTR is not associated with the antipsychotic-induced RLS in schizophrenia. To confirm these results, larger-scale association study is needed in the future.

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Mechanical Properties of Recycled Powder mixing Concrete (재생미분말을 혼입한 콘크리트의 역학적 특성)

  • Lee, Seung-Hwan;Jung, Dae-Jin;Choi, Ik-Chang
    • Proceedings of the Korea Concrete Institute Conference
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    • 2008.04a
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    • pp.769-772
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    • 2008
  • The problem of disposing construction waste materials has become a national and social problem. Recycled powder generated in the process of making aggregate, and the recycled powder is land-filled in its entirety. Results of toxicity testing of recycled power show that it contains base-pair substituent mutagenicity. As recycled powder is disposed of as landfill, it can cause secondary contamination such as soil and underground water contamination. There has been very little research made on recycled powder. This study has examined the utilization of concrete mixture by using recycled powder in a mixture instead of cement and compared and analyzed the characteristics of dynamics and workability. This study has examined the application of recycled powder in concrete. Depending on the replacement rate and workability, test piece was manufactured using different mixing rate by CP, WCP, PCP. The CP was used to examine the physical property of concrete and characteristics its dynamics. The letters W of WCP and P of PCP are the initials of water and mixture. They were made using the standard mixing ratiosemphasizing the workability to determine the characteristic of dynamics of concrete based on the mixing ratio of recycled powder. With the increase in the replacement rate, CP had very little change in the strength. But with the decline of slump, the workability was not good. The result of manufacturing WCP and PCP using the standard mixing ratio showed that WCP had a drop in strength compared to the plain. PCP had almost the same value as the plain only when the replacement rate was 10%. When it was higher than that, a reduction in strength was observed.

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Termination of mutant T7 RNA polymerases on intrinsic hairpin-independent termination signal (돌연변이 T7 RNA 증합효소의 머리핀 구조가 만들어지지 않는 인자독립형 전사종결 부위에서의 전사종결에 관한 연구)

  • Shin, Ji-Young;Kim, Dong-Hui;Lee, Sang-Soo
    • The Journal of Natural Sciences
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    • v.14 no.1
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    • pp.25-37
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    • 2004
  • T7 RNA polymerase also recognize intrinsic, hairpin-independent termination signal, a conserved 7-base pair sequence (ATCTGTT in the non-template stand) and U-rich sequence downstream of it. These intrinsic, hairpin-independent termination signal were commonly found in PTH and CJ termination sequences. There are two types of mutant T7 RNA polymerases recognizing sensitively(X3, X19, BG8) of insensitively (R173C) the intrinsic termination signals. We determined the T7 transcription activities of these mutants. Compared to wild-type, mutants X4, 19 and BG8 show highly reduced transcription activities (8%, 33%, 34%). On the other hand mutant R173C shows comparable transcription activity of wild-type (112%). Also transcription termination efficiencies at the PTH or CJ termination signals were determined by using mutant RNA polymerases. Temination of mutnats X4, X19 and BG8 are increased compared to wild-type. On the other hadn mutant R173C proceeds through PTH and CJ termination signals.

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Identification of Mycobacterium tuberculosis in Pleural Effusion by Polymerase Chain Reaction(PCR) (흉막 삼출액에서 중합효소 연쇄반응(PCR)을 이용한 M. tuberculosis의 검출)

  • Kim, Sun-Taec;Gang, Chang Woon
    • Tuberculosis and Respiratory Diseases
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    • v.42 no.5
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    • pp.695-702
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    • 1995
  • Background: Since polymerase chain reaction(PCR) was devised by Saiki in 1985, it has been used extensively in various fields of molecular biology. Clinically, PCR is especially useful in situation when microbiological or serological diagnosis is limited by scanty amount of causative agents. Thus, PCR can provide rapid and sensitive way of detecting M. tuberculosis in tuberculosis pleurisy which is diagnosed in only about 60 % of cases by conventional method. Method: To evaluate the diagnostic usefulness of PCR in tuberculosis pleurisy, The results of PCR was compared with those of conventional method, including pleural biopsy. The pleural effusion fluid was collected from 7 proven patients, 7 clinically suspected patients and control group(7 patients with malignant effusion). We extracted DNA from pleural fluid by modified method of Eisennach method(1991). The amplification target for PCR was 123 base pair DNA, a part of IS6110. Result: 1) Sensitivity of PCR: We detected upto 50fg DNA. 2) In patients with pleural effusion of proven tuberculosis, the positive rate of PCR was 85.7%(6/7). In patients with pleural effusion of clinically suspected tuberculosis, the positive rate was 71.5%(5/7). In control group, positive rate was 0%(0/7). Conclusion: We concluded that PCR method could be a very rapid, sensitive and specific one for diagnosis of M tuberculosis in pleural effusion. Further studies should be followed for the development of easier method.

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Construction and Characterization of a cDNA Library from the Camelina sativa L. as an Alternative Oil-Seed Crop (신 바이오디젤 원료 작물인 Camelina의 cDNA library 제작 및 유전자 특성)

  • Park, Won;Jang, Young-Seok;Ahn, Sung-Ju
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.55 no.2
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    • pp.151-158
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    • 2010
  • Camelina sativa L., known as popular names "gold-of-pleasure" or "false flax" is an alternative oilseed crop that can be grown under different climatic and soil conditions. Up to date, however, the genomic information of Camelina has not been studied in detail. Therefore, a cDNA library was constructed and characterized from young leaves. The constructed cDNA library incorporated of 1334 cDNA clones and the size of the insertion fragments average was 736 base pair. We generated a total of 1269 high-quality expressed sequence tags (ESTs) sequences. The result of cluster analysis of EST sequences showed that the number of unigene was 851. According to subsequent analysis, the 476 (55.9%) unigenes were highly homologous to known function genes and the other 375 (44.1%) unigenes were unknown. Remaining 63 (7.4%) unigenes had no homology with any other peptide in NCBI database, indicating that these seemed to be novel genes expressed in leaves of Camelina. The database-matched ESTs were further classified into 17 categories according to their functional annotation. The most abundant of categories were "protein with binding function or cofactor requirement (27%)", "metabolism (11%)", "subcellular localization (11%)", "cellular transport, transport facilities and transport routes (7%)", "energy (6%)", "regulation of metabolism and protein function (6%)". Our result in this study provides an overview of mRNA expression profile and a basal genetic information of Camelina as an oilseed crop.

Ansanella granifera gen. et sp. nov. (Dinophyceae), a new dinoflagellate from the coastal waters of Korea

  • Jeong, Hae Jin;Jang, Se Hyeon;Moestrup, Ojvind;Kang, Nam Seon;Lee, Sung Yeon;Potvin, Eric;Noh, Jae Hoon
    • ALGAE
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    • v.29 no.2
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    • pp.75-99
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    • 2014
  • A small dinoflagellate, Ansanella granifera gen. et sp. nov., was isolated from estuarine and marine waters, and examined by light microscopy, scanning electron microscopy, and transmission electron microscopy. In addition, the identity of the sequences (3,663-bp product) of the small subunit (SSU), internal transcribed spacer (ITS) region (ITS1, 5.8S, ITS2), and D1-D3 large subunit (LSU) rDNA were determined. This newly isolated, thin-walled dinoflagellate has a type E eyespot and a single elongated apical vesicle, and it is closely related to species belonging to the family Suessiaceae. A. granifera has 10-14 horizontal rows of amphiesmal vesicles, comparable to Biecheleria spp. and Biecheleriopsis adriatica, but greater in number than in other species of the family Suessiaceae. Unlike Biecheleria spp. and B. adriatica, A. granifera has grana-like thylakoids. Further, A. granifera lacks a nuclear fibrous connective, which is present in B. adriatica. B. adriatica and A. granifera also show a morphological difference in the shape of the margin of the cingulum. In A. granifera, the cingular margin formed a zigzag line, and in B. adriatica a straight line, especially on the dorsal side of the cell. The episome is conical with a round apex, whereas the hyposome is trapezoidal. Cells growing photosynthetically are $10.0-15.0{\mu}m$ long and $8.5-12.4{\mu}m$ wide. The cingulum is descending, the two ends displaced about its own width. Cells of A. granifera contain 5-8 peripheral chloroplasts, stalked pyrenoids, and a pusule system, but lack nuclear envelope chambers, a nuclear fibrous connective, lamellar body, rhizocysts, and a peduncle. The main accessory pigment is peridinin. The SSU, ITS regions, and D1-D3 LSU rDNA sequences differ by 1.2-7.4%, >8.8%, and >2.5%, respectively, from those of the other known genera in the order Suessiales. Moreover, the SSU rDNA sequence differed by 1-2% from that of the three most closely related species, Polarella glacialis, Pelagodinium bei, and Protodinium simplex. In addition, the ITS1-5.8S-ITS2 rDNA sequence differed by 16-19% from that of the three most closely related species, Gymnodinium corii, Pr. simplex, and Pel. bei, and the LSU rDNA sequence differed by 3-4% from that of the three most closely related species, Protodinium sp. CCMP419, B. adriatica, and Gymnodinium sp. CCMP425. A. granifera had a 51-base pair fragment in domain D2 of the large subunit of ribosomal DNA, which is absent in the genus Biecheleria. In the phylogenetic tree based on the SSU and LSU sequences, A. granifera is located in the large clade of the family Suessiaceae, but it forms an independent clade.

IDENTIFICATION OF PORPHYROMONAS ENDODONTALIS USING POLYMERASE CHAIN REACTION(RCR) (중합효소연쇄반응(Polymerase Chain Reaction)을 이용한 Porphyromonas endodontalis의 동정에 대한 연구)

  • Lee, Sang-Yup;Yoon, Soo-Han
    • Restorative Dentistry and Endodontics
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    • v.23 no.1
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    • pp.328-338
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    • 1998
  • Porphyromonas endodontalis, an anaerobic Gram negative cocobacillus which was known to be associated with the infected root canals and periapical lesions, is very difficult to culture and to detect by the traditional method in that it requires much time to induce the specific black pigmentation, and it is very sensitive to oxygen and the antibiotics added in the culture medium. In this study, the nucleotide sequences of the 'probe h' (0.73kb), one of the specific DNA probes top. endodontalis (ATCC 35406) which had been developed by our department, was determined and then a pair of primers for PCR amplification was fabricated to identify P. endodontalis. The plasmids containing 'probe h' were purified by $Wizard^{TM}$ Midipreps DNA Purification System (Promega Corp.), and the nucleotide sequences of the 'probe h' were determined by the dideoxy chain termination method using TaqTrack Sequencing System (Promega Corp.) and detected by fluorescent labelling method. The sense/antisense PCR primers were designed with computer software (Lasergene, DNASTAR Ind. PCR was done with a programmable GeneAmp PCR System 2400 (Perkin Elmer-Cetus Co.). Each sample containing the whole genomic DNA of P. endodontalis and other black-pigmented Bacteroides was itailly denatured at $94^{\circ}C$ for 5 min and then subjected to 30 cycles, each of them consisting of 60s at $94^{\circ}C$, 60s at $60^{\circ}C$, and 90s. at $72^{\circ}C$. The amplified DNA was resolved electrophoretically in a 1.0 % agarose gel in 1X TAE buffer, stained with EtBr, and photographed on a UV transilluminator. The results were as follows : 1. The nucleotide sequences of 'probe h' (743 base pairs) were obtained by dideoxy chain termination method, and from that results the specific primers to P. endodontalis (ATCC 35406), 'Primer H1/ Primer H2', were designed. 2. It has been found that 'Primer H1/H2' could detect P. endodontalis (ATCC 35406) using PCR. 3. The PCR system with this primers may be a powerful technique to amplify the specific sequences of 'probe h' of P. endodontalis (ATCC 35406) that produce distinct identification of it from other black-pigmented Bacteroides, and this could help us to determine the nature of periapical disease.

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Development of SSLP Marker Targeted to P34 Null Gene in Soybean (콩 P34 단백질 결핍 유전자를 이용한 SSLP 마커 개발)

  • Yang, Kiwoung;Ko, Jong-Min;Lee, Young-Hoon;Jeon, Myeong Gi;Jung, Chan-Sik;Baek, In-Youl;Kim, Hyun-Tae;Park, Keum-Yong
    • Korean Journal of Breeding Science
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    • v.42 no.5
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    • pp.502-506
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    • 2010
  • Soybean seed possesses about 15 allergenic proteins recognized by IgEs from soy-sensitive human. The allergenic impact of soybean proteins limit its extensive usage in a broad range of processed foods. Soybean protein P34 or Gly m Bd 30k of the cysteine protease family is one of the major allergen of the soybean seed. P34-null soybean, PI567476, was identified among soybean (Glycine max & Glycine soja Sieb. and Zucc) of approximately 16,226 accessions from USDA soybean germplasm screened. Also, for P34 gene (Williams 82; whole genome sequence cultivar) and P34 null gene (PI567476) comparative analysis of sequences listed in the NCBI database showed the presence of a SSLP (Simple Sequence Length Polymorphism) of 4 base pair. So, a SSLP marker was designed to reveal the polymorphism of the locus. In this study, a population of 339 $F_2$ recombinant inbred lines generated by cross between Taekwang (Glycine max) and PI567476 was used to select $F_{2:3}$ plant of a P34 null gene. The result separation rate Taekwang type, heterozygous type and PI567476 type were shown in 85: 187: 67 since single gene is concerned in as the separation rate of 1:2:1 in $X^2{_{0.05}}=5.99$, df=2. In future, selected plant will identify protein level, whether P34 null protein is equal to P34 null gene.

Bird accidents in Southern Mongolia: a case study of bird electrocution (몽골 남부지역의 야생조류 사고: 감전사를 중심으로)

  • Ganbold, Onolragchaa;Bing, Gi-Chang;Purevee, Erdenetushig;Munkhbayar, Munkhbaatar;Choi, Won-Suk;Jargalsaikhan, Ariunbold;Paik, In-Hwan;Purevdorj, Zoljargal;Jargal, Namsrai;Paek, Woon-Kee
    • Korean Journal of Ornithology
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    • v.25 no.2
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    • pp.94-100
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    • 2018
  • Bird electrocution on contact with electricity lines is well reported and is quite common in Mongolian open areas. We visited 15-kV electricity distribution pole lines in a Southern Mongolian semi-desert region three times in 2017, in April, July, and September, to assess their risks to birds. The carcasses of 45 electrocuted birds representing 12 species were identified from 250 poles (overall mortality rate of 1.12% every 10 km). The majority of these electrocuted birds were endangered Saker Falcon (n=11) and common Black Kite (n=11). The lacks of roosting or perching sites in our study sites (indeed other open areas in Mongolia) may resulted for such high rate bird electrocution. A 751-base pair (bp) cytochrome c oxidase I (COI) gene was amplified for DNA sequence-based identification of carcasses that were difficult to identify directly. Our findings revealed the high electrocution risk for birds in Mongolian open areas, especially for the birds of prey, with relatively low-efficiency electrocution mitigation approaches. The findings also indicate that there is a need for better understanding of the risk of bird electrocution, particularly in the open areas. This will contribute to the conservation of endangered species.