• 제목/요약/키워드: rDNAs

검색결과 171건 처리시간 0.025초

Cloning, Expression and Genomic Organization of Genes Encoding Major Royal Jelly Protein 1 and 2 of the Honey Bee (Apis cerana)

  • Imjongjirak, Chanprapa;Klinbunga, Sirawut;Sittipraneed, Siriporn
    • BMB Reports
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    • 제38권1호
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    • pp.49-57
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    • 2005
  • Major Royal Jelly Protein cDNAs of Apis cerana (AcMRJP) were cloned and characterized. The open reading frames (ORFs) of the AcMRJP1 and AcMRJP2 genes were 1302 and 1392 nucleotides, encoding 433 and 463 amino acid residues, respectively. The sequence divergences between AcMRJP1 and AcMRJP2 and their corresponding protein families in A. mellifera were 0.0618 and 0.0934 at the nucleotide level and 0.0912 and 0.1438 at the protein level, respectively. Phylogenetic analysis supports the orthologous similarity between these proteins. The deduced amino acids indicated high essential amino acid contents of AcMRJP1 and AcMRJP2 (47.5 and 44.8%, respectively). The genomic organization of both AcMRJP1 and AcMRJP2 was determined. Both the AcMRJP1 (3663 bp) and AcMRJP2 (3963 bp) genes contained six exons and five introns, where all boundaries conformed to the GT/AG rule. AcMRJP1 and AcMRJP2 cDNAs were cloned into pET17b, and both the recombinant (r) AcMRJP1 (47.9 kDa) and rAcMRJP2 (51.7 kDa) were expressed in the insoluble form. Western blot analysis and N-terminal sequencing of the solubilized proteins revealed successful expression of rAcMRJP1 and rAcMRJP2 in vitro. The yields of the purified rAcMRJP1 and rAcMRJP2 were approximately 20 and 8mg protein per liter of the flask culture, respectively.

Cytogenetic Mapping of Carthamus tinctorius L. with Tandemly Repeated DNA Sequences by Fluorescence in situ Hybridization

  • Mancia, Franklin Hinosa;Ju, Yoon Ha;Lim, Ki-Byung;Kim, Jung Sun;Nam, Sang Yong;Hwang, Yoon-Jung
    • 한국자원식물학회지
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    • 제30권6호
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    • pp.654-661
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    • 2017
  • Dual-color fluorescence in situ hybridization karyotype analysis was created using repetitive sequences including two types of rDNA repeats (45S and 5S rDNAs) and Arabidopsis-type telomere sequence repeats. The somatic metaphase cells of Carthamus tinctorius were observed as diploids (2n=2x=24). A symmetrical or slightly asymmetrical karyotype with seven pairs of metacentric and five pairs of submetacentric chromosomes was observed. The lengths of the somatic metaphase chromosomes ranged from 4.18 to $6.53{\mu}m$, with a total length of $60.71{\mu}m$. One locus of 45S rDNA was located on the pericentromeric regions of three pairs of chromosomes and the other pair was situated on the terminal regions of the short arms of a single pair of chromosomes. One locus of 5S rDNA was detected on the interstitial regions of the short arms of two pairs of chromosomes. Arabidopsis-type telomeric repeats were detected on the terminal regions of all pairs of chromosomes. Co-localization of loci between telomeric repeats and 45S rDNA was observed in a single pair of chromosomes. The results provide additional information for the existing physical mapping project of C. tinctorius and will also serve as a benchmark to a more intricate cytogenetic investigation of C. tinctorius and its related species.

Genetic Analysis of Haimen Chicken Populations Using Decamer Random Markers

  • Olowofeso, O.;Wang, J.Y.;Zhang, P.;Dai, G.J.;Sheng, H.W.;Wu, R.;Wu, X.
    • Asian-Australasian Journal of Animal Sciences
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    • 제19권11호
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    • pp.1519-1523
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    • 2006
  • Through a screening and selection approach method, decamer random markers were used in a technique called random amplified polymorphic DNA (RAPD) assay with 252 genomic DNAs isolated from four major Haimen chicken populations: Rugao (62), Jiangchun (62), Wan-Nan (63) and Cshiqishi (65). A total of 3-score decamer random primers (S241-S260, S1081-S1100 and S1341-S1360) were employed in the preliminary RAPD-polymerase chain reaction (RAPD-PCR) assay with 50 random template DNA samples from all the populations. Four (6.67%) of the primers that produced obvious polymorphic patterns, interpretable and reproducible bands were selected and used with both the individual DNAs from each population and with pooled DNA samples of the four populations in subsequent analyses. The selected primers produced a total of 131 fragments with molecular size ranging from 835 to 4,972 base pairs (bp) when used with the individual DNAs; 105 (80.15%) of these fragments were polymorphic. With the pooled DNAs, 47 stable and characteristic bands with molecular size ranging from 840 to 4,983 bp, of which 23 (48.94%) polymorphic, were also generated. The band-sharing coefficient (BSC) calculated for the individuals in the population and among populations of bulked samples was between 0.8247 (Rugao) and 0.9500 (Cshiqishi); for pairwise populations, it was between 0.7273 (Rugao vs. Wan-Nan) and 0.9367 (Jiangchun vs. Cshiqishi) chicken populations. Using the BSC for individual and pairwise populations, the Nei's standard genetic distances between the chicken populations were determined and ranged from 0.0043 (Jiangchun vs. Cshiqishi) to 0.1375 (Rugao vs. Cshiqishi). The reconstructed dendrogram linked the Jiangchun and Cshiqishi chickens as closely related populations, followed by Wan-Nan, while the Rugao was the most genetically distant among the populations.

Application of Chromosome Manipulation, DOP-PCR and AFLP Methods to Isolate Sex-Specific DNAs from Rumex acetosa L.

  • Jin, Dong-Chung;Kim, Joong-Soon;Park, ji-Young;Bong, Jae-Wook;Hur, Yoon-Kang
    • Journal of Photoscience
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    • 제12권2호
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    • pp.75-82
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    • 2005
  • Rumex acetosa L. is a dioecious flowering plant with well developed sex chromosome system: 2n = 12 + XX in the female plants and 2n = 12 + XY1Y2 in the male plants. To isolate sex-linked DNA, we carried out chromosome micromanipulation, followed by DOP-PCR, AFLP of the PCR products, reverse Southern hybridization and sequence analysis. From 500 AFLP specific clones, 13 X-chromosome and 5 Y-chromosome specific clones were obtained. Except one clone RADAX-239 ($\underline{R}umex\;\underline{a}-\underline{D}OP-PCR-\underline{A}FLP-\underline{Y}-chromosome\;specific$), all clones appear to be R. acetosa plant-specific sequences and non-coding sequences. Southern blot analysis using these clones could not discriminate genomic DNAs either from male or female plants. Results of this study imply that both autosome-origin and degeneration of sex chromosomes are prevalent in plant systems.

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Morphological and genetic diversity of Euglena deses group (Euglenophyceae) with emphasis on cryptic species

  • Kim, Jong Im;Linton, Eric W.;Shin, Woongghi
    • ALGAE
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    • 제31권3호
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    • pp.219-230
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    • 2016
  • The Euglena deses group are common freshwater species composed of E. adhaerens, E. carterae, E. deses, E. mutabilis, and E. satelles. These species are characterized by elongated cylindrical worm-like cell bodies and numerous discoid chloroplasts with a naked pyrenoid. To understand the cryptic diversity, species delimitation and phylogenetic relationships among members of the group, we analyzed morphological data (light and scanning electron microscopy) and molecular data (nuclear small subunit [SSU] and large subunit [LSU] rDNAs and plastid SSU and LSU rDNAs). Bayesian and maximum likelihood analyses based on the combined four-gene dataset resulted in a tree consisting of two major clades within the group. The first clade was composed of two subclades: the E. mutabilis subclade, and the E. satelles, E. carterae, and E. adhaerens subclade. The E. mutabilis subclade was characterized by a lateral canal opening at the anterior end and a single pellicular stria, whereas the E. satelles, E. carterae, and E. adhaerens subclade was characterized by an apical canal opening at the anterior end of the cell and double pellicular striae. The second clade consisted of 20 strains of E. deses, characterizing by a subapical canal opening at the anterior end and double pellicular striae, but they showed cell size variation and high genetic diversity. Species boundaries were tested using a Bayesian multi-locus species delimitation method, resulting in the recognition of five cryptic species within E. deses clade.

울릉도 연안 수심 1500 m에 서식하는 해양미생물군집의 분포 (Marine Prokaryotic Diversity of the Deep Sea Waters at the Depth of 1500 m Off the Coast of the Ulleung Island in the East Sea (Korea))

  • 김미경;강용호
    • 미생물학회지
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    • 제48권4호
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    • pp.328-331
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    • 2012
  • 울릉도 연안의 심해(1500 m)에 서식하는 미생물의 다양성을 조사하였다. Ultramembrane filter를 사용하여 해양미생물을 여과한 다음 미생물군집 DNA를 정제하여 16S rDNA를 증폭하였다. Pyrosequencing 방법으로 염기서열을 분석한 결과 총 13,029 reads를 얻었으며, 이중에 54.1%가 uncultured bacteria, 23.4%가 alphaproteobacteria, 22.3%가 gammaproteobacteria이었고 flavobacteria, actinobacteria, epsilonproteobacteria 등이 0.2%이내에서 분포하고 있었다. 울릉도 지역의 해양심층수에서 배양이 가능한 것으로 알려진 미생물로서는 alphaproteobacteria의 Rhodobacteraceae과 (family), gammaproteobacteria의 Alteromonadaceae, Halomonadaceae, Piscirickettsiaceae과가 주로 분포하였다.

Distribution Patterns of the Members of Phylum Acidobacteria in Global Soil Samples

  • Lee, Sang-Hoon;Cho, Jae-Chang
    • Journal of Microbiology and Biotechnology
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    • 제19권11호
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    • pp.1281-1287
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    • 2009
  • The distribution pattern of the phylum Acidobacteria, a previously uncultured bacterial group, was investigated by molecular ecological analyses of global soil samples collected from pristine ecosystems across five continents. Acidobacterial 16S rDNAs were observed in almost all soil samples, and members of acidobacterial primer group A were detected in all samples that harbored the phylum Acidobacteria. Other primer groups, Y, G, and O, showed limited distribution patterns. We further divided the primer groups into acidobacterial subdivisions (class-level). Subdivisional distribution patterns were determined by comparing the observed T-RFs with theoretical T-RFs predicted by in silico digestion of acidobacterial 16S rDNAs. Consistent with the PCR results obtained with subgroup-specific primers, T-RFLP analyses showed that acidobacterial subdivision 1 belonging to primer group A was present in the majority of the soil samples. This study revealed that the phylum Acidobacteria could be globally distributed. At the subdivisional level, acidobacterial subdivision 1 might be the most widely distributed group in this phylum, indicating that members of subdivision 1 might be adapted to various soil environments, and members belonging to other subdivisions might be restricted to certain geographic regions or habitats.

Rhizina undulata rDNA ITS 영역의 PCR 검정 및 염기배열 분석 (PCR Detection and Sequence Analysis of the rDNA ITS Regions of Rhizina undulata)

  • 이선근;이종규;김경희;이승규;이상용
    • 한국산림과학회지
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    • 제96권4호
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    • pp.425-431
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    • 2007
  • Rhizina undulata의 PCR 검정 및 유전적 특성 분석을 목적으로, rDNA ITS 영역의 염기배열 해석 및 PCR 방법에 의한 토양으로부터 R. undulata의 진단법을 개발하였다. 18S rDNA 부분의 염기서열 분석 결과, 공시한 4종의 균주 모두 1,375 nt의 크기로 동일하였으며, 염기배열도 100% 일치하였다. 한편, rDNA ITS 영역의 염기배열은 585 nt이었고, PDK-1, PTT-1 및 PDJ-9 균주는 염기배열이 100% 동일하였으나, PDS-5균주에서는 두 곳에서 염기의 치환이 발견되었다. 이와 같은 염기배열을 분석하여 제작한 R. undulata rDNA ITS 영역 특이적 primer를 이용한 PCR 검정 결과, R. undulata 균주들에서만 약 525 bp 크기의 ITS 영역 특이적인 증폭산물이 검출되었다. PCR 방법에 의하여 검출할 수 있는 토양 중의 R. undulata 최소 균사량의 한계를 확인하기 위해서, 순수 배양한 R. undulata 균사현탁액을 순차 희석하여 100g의 사양토에 혼합한 다음, 농도별로 균사 혼합한 각각의 토양 시료로부터 추출한 total DNA의 PCR 증폭산물을 분석한 결과, PCR 방법에 의하여 100g의 토양 중에 1 ng의 R. undulata 균사가 함유되어 있는 경우까지 검출이 가능하였다.