• Title/Summary/Keyword: 185 rDNA

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Morphology and Sequence Analysis of Nuclear 18S rDNA from the Summer Strain of Porphyra suborbiculata (Rhodophyta) in Korea (여름철 서식 한국산 홍조류 둥근돌김 (Porphyra suborbiculata)의 형태 및 18S rDNA 염기서열 분석)

  • JIN Long-Guo;KIM Myung-Sook;CHOI Jae-Suk;CHO Ji-Young;JIN Hyung-Joo;HONG Yong-Ki
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.33 no.6
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    • pp.489-495
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    • 2000
  • The 185 ribosomal RNA gene (185 rDNA) of the marine alga Porphyra sp. 723 (Bangiales, Rhodophyta) was amplified using the polymerase chain reaction and its sequence was analysed. The Porphyra species was a summer strain collected on rocks in upper intertidal zone at Ikidae, Pusan on 23rd July 1999. The fronds were $1{\~}5 cm$ long, monostromatic, and orbicular or ovate shaped, They had spinulate processes at margin of the frond, Comparison of this 185 rDNA sequence with the other Forphyra species indicates that Porphyra sp. 723 has the same 185 rDNA sequence derived from Porphyra suborbiculata (NCBI access number; AB 013180) except one base pair substitution in 2327 base pairs.

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Sequence Analysis of Nuclear 18S rDNA from Porphya tenera (Rhodophyta) in Korea (한국산 참김 (Porphya tenera)의 핵 18S rDNA염기서열 분석)

  • JIN Long-Guo;KIM Myung-Sook;CHOI Jae-Suk;CHO Ji-Young;JIN Duck-Hee;HONG Yong-Ki
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.36 no.1
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    • pp.35-38
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    • 2003
  • Nuclear 18S ribosomal RNA gene (185 rDNA) from the aquaculturable seaweed Porphya tenera (Bangiales, Rhodophyta) was amplified using the polymerase chain reaction and its sequence was analysed. Complete 185 rDNA has an 1,822 bp exon and a 510 bp intron. The G+C contents of exon and intron were $48.68\%\;and\;54,90\%,$ respectively. The exon sequence showed $99.6\%$ homology to the GebBank accession number AB029880 of the Japanese P. tenera. The intron region that is inserted upstream between 568 and 1,079 showed $43.6\%$ homology to the AB029880.

Sequence Analysis of Nuclear 18S rDNA from the Seaweed Porphyra yezoensis (Rhodophyta) in Korea (한국산 방사무의김 (Porphyra yezoensis)의 핵 18S rDNA 염기서열 분석)

  • Jin Long Guo;Kim Myung Sook;Choi Jae Suk;Cho Ji Young;Jin Hyung Joo;Hong Yong Ki
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.35 no.6
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    • pp.633-638
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    • 2002
  • Nuclear 18S ribosomal RNA gene (185 rDNA) from the aquaculturable seaweed Porphya yezoensis (Bangiales, Rhodophyta) was amplified using the polymerase chain reaction and its sequence was analysed. Complete 185 rDNA has an 1823 bp exon and a 514 bp intron. The G+ C contents of exon and intron were $48\%$ and $51.4\%$, respectively. The exon sequence showed $99.5\%$ homology to the GenBank accession number AB013177 of the Japanese p. yezoensis. The intron region that was inserted upstream between 568 and 1083 showed $93.4\%$ homology to the AB013177.

Sequence Similarity of Nuclear 18S rDNA from Morphologically Different Blades of the Seaweed Porphyra pseudolinearis (Rhodophyta) (긴잎돌김 Porphyra pseudolinearis의 엽체형간 18S rDNA 염기서열 상동성)

  • Jin Long-Guo;KIM Young-Dae;KIM Myung-Sook;JIN Hyung-Joo;CHO Ji-Young;CHOI Jae-Suk;HONG Yong-Ki;KIM Hyung Geun
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.33 no.6
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    • pp.496-500
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    • 2000
  • Partial fragments of nuclear 185 rDNAs from morphologically wide and narrow thalli of the seaweed Porphyra pseudolineazis were amplified and sequenced to compare their DNA homology. Both sequences of 311 base pairs showed $100{\%}$ identical each other. They showed $97.7{\%}$ similarity with a wild strain collected at Sodol in Kangwondo, and $99.4{\%}$ similarity with the GenBank accession number AB013185 of the Japanese P. pseudolinearis. Thus the morphological difference of wide and narrow blades might not be a classification criterion for the sub-species level of P. pseudolinearis.

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Clonorchis sinensis tropomyosin: Cloning and sequence of partial cDNA amplified by PCR (간흡충 tropomyosin: PCR로 일부분 증폭된 cDNA의 cloning 및 염기서열)

  • 홍성종
    • Parasites, Hosts and Diseases
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    • v.31 no.3
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    • pp.285-292
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    • 1993
  • C. sinensis total RMh was containing large amount of 185 rRNA but little 285 rRNA. The size of the double-stranded cDNA synthesized from poly $(A)^{+}$ mRNA was 0.4-4.2 kb long with tapering unto 9.5 kb. Degenerated oligonucleotides (as 2 sense and 3 antisense Primers) were designed on the conserved regions of the known tropomyosin amino acid sequences. From one out of the PCR amplifications using total CDNA and matrix of primers, a specific gene product, 580 bp in size, was produced. Upon Southern hybridization of the PCR products with Schistosomn mnnsoni tropomyosin (SMTM) CDNA, only one signal appeared at the band of 580 bp product. This 580 bp product was considered to encode C. sinensis tropomyosin (CSTM) and cloned in pGEM-3Zf(-) for DNA sequencing. CSTM cDNA was 575 bp containing one open reading frame of 191 predicted amino acids, which revealed 86.3% homology with SMTM and 51.1% with rrichostronsylur coeubnlormis tropomyosin. CSTM cDNA obtained will serve as a probe in the studies of molecular cloning of CSTM.

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Sequence Analysis of Nuclear 18s rDNA from Porphyra dentata (Rhodophyta) in Korea (한국산 잇바디돌김 (Porphyra dentata)의 핵 18S rDNA 염기선열 분석)

  • Long-Guo Jin
    • Journal of Life Science
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    • v.12 no.4
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    • pp.427-432
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    • 2002
  • Nuclear 18S ribosomal RNA gene (18S rDNA or SSU rDNA) from the Porphyra dentata tissue was amplified and sequenced. Complete 18S rDNA has an 1822 bp exon and a 512 bp intron. The G+C contents of exon and intron were 49% and 55%, respectively. The exon sequence showed 97.1% homology to the GenBank accession number AB013183 of the Japanese P. dentata. The intron region that is inserted in upstream between 568 and 569 showed 52.1% homology to the AB013183.

Species Identification of Five Penaeid Shrimps Using PCR-RFLP and SSCP Analyses of 16S Ribosomal DNA

  • Khamnamtong, Bavornlak;Klinbunga, Sirawut;Menasveta, Piamsak
    • BMB Reports
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    • v.38 no.4
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    • pp.491-499
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    • 2005
  • DNA-based molecular markers for differentiation of five penaeid shrimps (Penaeus monodon, P. semisulcatus, Feneropenaeus merguiensis, Litopenaeus vannamei and Marsupenaeus japonicus) were developed based on polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and single-stranded conformation polymorphism (SSCP) of 16S ribosomal (r) DNA. Differentiation of P. monodon, P. semisulcatus and L. vannamei can be unambiguously carried out by PCR-RFLP of 16S $rDNA_{560}$ whereas P. semisulcatus and M. japonicus shared a BABB mitotype. These shrimps were successfully discriminated by SSCP analysis of 16S $rDNA_{560}$. Nevertheless, the amplification success for L. vannamei and F. merguiensis was not consistent when tested against larger sample sizes. As a result, 16S $rDNA_{560}$ of an individual representing the most common mitotype of each species was cloned and sequenced. The new primer pair was designed and tested against the large sample sizes (312 bp product, N = 185). The amplification success was consistent across all species. PCR-RFLP of 16S $rDNA_{312}$ was as effective as that of 16S $rDNA_{560}$. Differentiation of all shrimp species were successfully carried out by SSCP analysis.

PCR of Gut Contents for a Food Web Study of a Marine Ecosystem

  • Kim, Nack-Keun;Kim, Kyoung-Sun;Kim, Hyun-Woo
    • Fisheries and Aquatic Sciences
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    • v.10 no.4
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    • pp.179-185
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    • 2007
  • Understanding dietary habits is one of the most important factors in studying food webs and other ecological processes. Here we designed universal primers to amplify portions of the 18S and 28S rDNA sequences to examine gut contents using PCR techniques. The gut contents of sailfin sandfish (Arctoscopus japonicus) and pacific squid (Todarodes pacificus) were examined. In total, 11 families of prey were identified with 18S and 28S rDNA using the universal primers. The DNA sequence data indicated that the primer sets successfully amplified a wide spectrum of species and represented gut contents in a relatively convenient way. We found that information in the NCBI database was not yet sufficient to discriminate the species we isolated. In addition, technology for the separation of heterogeneous PCR products and better resolution and quantification protocols would help increase data accuracy.

Phylogeny of Mite Taxa (Acari : Sarcoptiformes) Based on Small Subunit Ribosomal RNA Sequences (리보솜 Small unit RNA 염기서열을 이용한 진드기류(Acari:Sarcoptiformes)의 분류)

  • Lee Keun Hee;Yu Hak Sun;Park Sang Kyun;Lee Sun Joo;Lee Kyeong Ah;Kim Sun Mee;Ock Mee Sun;Jeong Hae Jin
    • Journal of Life Science
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    • v.16 no.1
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    • pp.71-75
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    • 2006
  • We analyzed the phylogenic relationships of 23 partial 18S rDNA sequences of 22 species (1 species has 2 strains) belonging to Sarcorptiforms include 4 new sequences, using several tools. Although geographic distributions are quite far from, sequence similarity of two strains of Dermatophygoides pteronyssinus isolated from Japan and New Zealand were very high. This result suggests that mite migration by animals including human occurred in the two continents. We investigated the Endeostigmata taxonomic relationship between the Prostigmata and Oribatida subgroups using small fragments (340-400 bp) of their 185 rDNA sequences. But Endeostigmata was not grouped with Oribatida or Prostigmata. In conclusion, it is first reported phylogenic relationship for classified mites included in Sarcoptiformes using 185 rDNA sequence analysis and its system is a very powerful tool for classification of mites.

Detection of Ralstonia solanacearum with Nested PCR and DNA Enzyme-Linked Immunosorbent Assay (Nested PCR과 DNA Enzyme-Linked Immunosorbent Assays를 이용한 Ralstonia solanacearum의 검출)

  • Ko, Young-Jin;Cho, Hong-Bum
    • Korean Journal of Microbiology
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    • v.43 no.3
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    • pp.179-185
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    • 2007
  • In this study, we used the method of guanidin isothiocyanate and boiling with Chelex-100 resin to extract genomic DNA of Ralstonia solanacearum from soil. It is more efficient than general protocols to remove inhibitory compounds in soil and R. solanacearum on. Then, we applied polymerase chain reaction and DNA enzyme-linked immunosorbent assay (ELISA) to identify and detect pathogen. The fliC gene of R. solanacearum was selected for specific detection of pathogen and primer sets were designed. Among the primer sets, two specific and sensitive primer sets, RsolfliC(forward: 5-GAACGCCAACGGTGCGAACT-3 and reverse; 5-GGCGGCCTTCAGGGAGGTC-3, designed by J. $Sch\ddot{o}nfeld$ et al.) and RS_247 (forward: 5-GGCGGTCTGTCGGCRG-3 and reverse; 5-CGGTCGCGTTGGCAAC-3 designed by this study), were designed to perform nested PCR. Nested PCR primer was labeled with biotin for hybridization between nested PCR product and probe to analyze with DNA ELISA.