• Title/Summary/Keyword: arbitrary primer

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Genetic Variation of the Wild Strains of Lentinula edodes in Three Mountains of Korea (계방산, 오대산 및 지리산 야생 표고균주의 유전적 변이)

  • Kim, Dool-Yi;Bak, Won-Chull
    • The Korean Journal of Mycology
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    • v.29 no.2
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    • pp.99-103
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    • 2001
  • Genetic variation of the wild strains of Lentinula edodes[(Berk.)Pegler] in three regions of Korea was investigated by analyzing random amplified polymorphic DNA (RAPD) markers. A total of 32 strains of L. edodes were collected from Mt. Kyebang (10 strains), Mt. Odae (11), and Mt. Jiri (11), respectively. The genomic DNA was amplified by polymerase chain reaction (PCR) using an arbitrary 10-mer primer. A total of 170 amplified fragments were observed, of which 161 fragments were polymorphic. The results of cluster analysis, performed on the basis of the presence or absence of amplified fragments of the same size, revealed that strains collected from both Mt. Kyebang and Mt. Odae in a single group. AMOVA analysis revealed that genetic variations between sites amounted to 12.5%, while 87.1% of total variations was explained by variations among strains within sites. Relatively high genetic relationships among the strains of Mt. Kyebang and Mt. Odae, which were high variance within populations. Whereas, all the strains of Mt. Jiri, which were low variance among populations from both Mt. Kyebang and Mt. Odae, which resulted in genetic isolation of the strains in Mt. Jiri.

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Isolation of an Acid-Labile Gene from the Seaweed Porphyra yezoensis Tissue (해조류 김 Porphyra yezoensis 엽체로부터 산에 민감한 유전자의 분리)

  • Jin, Hyung-Joo;Park, Sun-Mee;Kim, Long-Guo;Jin, Deuk-Hee;Kong, In-Soo;Hong, Yong-Ki
    • KSBB Journal
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    • v.14 no.6
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    • pp.702-706
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    • 1999
  • The genetic responses of aquaculturable seaweed Prophyra yezoensis tissue by acid shock have been compared using differential display technique. The tissue has been challenged in seawater containing 0.05% hydrogen chloride(pH 3.0) for 5 min, then rehabilitated in normal seawater for 10 min, 30 min, 60 min and 4 hrs, respectively. Total RNA was extracted by LiCl-guanidinium method. The cDNA was synthesized by reverse transcription with random hexamers and amplified by PCR with arbitrary primers. The genetic fragment disappeared by acid shock was selectively isolated from agarose gel and sequenced with a DNA auto sequencer. One of the acid-labile gene(605 bp) was identified as a dethiobiotin synthetase gene according to sequence alignment analysis by the NCBI BLAST search program.

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Genetic Relationships among the Poplars of Section Leuce (Genus Populus) revealed by RAPD Marker Analysis (RAPD 표식자(標識者) 분석(分析)에 의한 사시나무속(屬) Leuce절(節) 포플러의 유연관계(類緣關係))

  • Hong, Kyung-Nak;Hyun, Jung Oh;Hong, Yong Pyo
    • Journal of Korean Society of Forest Science
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    • v.87 no.2
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    • pp.153-163
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    • 1998
  • Genetic relationships of some poplars in the section Leuce, including 5 species and 11 clones of Populus alba${\times}$glandulosa, were investigated on the basis of RAPD marker analysis. Twenty-two of the 88 arbitrary 10-mer primers, showed reproducible amplification in the preliminary experiment with 6 samples, were used for PCR and generated a total of 181 RAPD markers. Genetic relationships among the analyzed samples were tested by two phenetic methods of the UPGMA and the neighbor-joining, which revealed the close genetic relationship between P. glandulosa and P. alba. And the close genetic relationship between P. glandulosa and P. davidiana was ascertained by the principal component analysis. Based on the observation of the close genetic relationship between them, it was deduced that P. glandulosa might be originated by the saltational speciation caused by the hybridization between P. alba and P. davidiana in nature.

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Phylogenetic relationships in different strains of Pholiota species based on PCR polymorphism (PCR 다형성 분석에 의한 비늘버섯 속 계통의 유연관계 분석)

  • Kwon, Woon-Hyuk;Park, Hyuk;Baek, Min-Jae;Cho, Woo-Jin;Choi, Woo-Jeong;Ahn, Chi-Beom;Shin, Do-Bin;Lee, Tae-Soo
    • Journal of Mushroom
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    • v.11 no.2
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    • pp.69-76
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    • 2013
  • Pholiota species were collected from different geographical regions of the world. Genetic diversity and phylogenetic relationships were analyzed by rDNA-ITS sequences and RAPD polymorphism. The sizes of rDNA-ITS PCR amplicons of Pholiota spp. varied from 233~271, 158~223 and 174~219 bp, respectively. A phylogenetic tree was constructed on the ITS region sequences and Pholiota strains were classified into 8 clusters. Twenty strains in seven Pholiota spp. were classified into seven clusters by RAPD polymorphism using 15 arbitrary primers. Our experimental results suggested that rDN-ITS and RAPD analysis are useful tool for classifying Pholiota spp. and strains.

Identification of Differentially Expressed Genes in the Longissimus Dorsi Muscle Tissue between Duroc and Erhualian Pigs by mRNA Differential Display

  • Pan, P.W.;Zhao, S.H.;Yu, M.;Liu, B.;Xiong, T.A.;Li, K.
    • Asian-Australasian Journal of Animal Sciences
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    • v.16 no.7
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    • pp.1066-1070
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    • 2003
  • In order to identify differentially expressed mRNAs (which represent possible candidates for significant phenotypic variances of muscle growth, meat quality between introduced European and Chinese indigenous pigs) in the longissimus dorsi muscle tissue between adult Duroc and Erhualian pigs, mRNA differential display was performed. Five 3' anchor primers in combination with 20 different 5' arbitrary primers (100 primer sets) were used and nearly 5,000 cDNA bands were examined, among which 10 differential display cDNAs were obtained, cloned and sequenced. Six of the 10 cDNAs showed similarity to identified genes from GenBank and the other 4 had no matches in GenBank. Differential expression was tested by Northern blot hybridization and could be confirmed for 2 cDNAs. The method used in this study provides a useful molecular tool to investigate genetic variation that occurs at the transcriptional level between different breeds.

Distinction between Cold-sensitive and -tolerant Jute by DNA Polymorphisms

  • Hossain, Mohammad Belayat;Awal, Aleya;Rahman, Mohammad Aminur;Haque, Samiul;Khan, Haseena
    • BMB Reports
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    • v.36 no.5
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    • pp.427-432
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    • 2003
  • Jute is the principal coarse fiber for commercial production and use in Bangladesh. Therefore, the development of a high-yielding and environmental-stress tolerant jute variety would be beneficial for the agro economy of Bangladesh. Two molecular fingerprinting techniques, random-amplified polymorphic DNA (RAPD) and amplified-fragment length polymorphism (AFLP) were applied on six jute samples. Two of them were cold-sensitive varieties and the remaining four were cold-tolerant accessions. RAPD and AFLP fingerprints were employed to generate polymorphism between the cold-sensitive varieties and cold-tolerant accessions because of their simplicity, and also because there is no available sequence information on jute. RAPD data were obtained by using 30 arbitrary oligonucleotide primers. Five primers were found to give polymorphism between the varieties that were tested. AFLP fingerprints were generated using 25 combinations of selective-amplification primers. Eight primer combinations gave the best results with 93 polymorphic fragments, and they were able to discriminate the two cold-sensitive and four cold-tolerant jute populations. A cluster analysis, based on the RAPD and AFLP fingerprint data, showed the population-specific grouping of individuals. This information could be useful later in marker-aided selection between the cold-sensitive varieties and cold-tolerant jute accessions.

Monitoring of Gentic Variability in Dicofol-susceptible, Dicofol-resistant, and its Reverse-selected Strains of Tetranychus urticae by RAPD-PCR

  • Song, Cheol;Park, Jin-Hee;Kim, Gil-Hah;Kwon, O-Yu;Cho, Kwang-Yun
    • Journal of Life Science
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    • v.9 no.1
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    • pp.14-16
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    • 1999
  • Genetic variability was monitored by random amplified polymorphic DNA-polymerase chain reaction (RAPD-PCR) in dicofol-susceptible (S), dicofol-resistant (R) and its reverse-selected (RS) strains of two-spotted spider mite, of Tetranychus urticae. Before the reverse-selection, RS strain, selected reversely from R strain, was 23-fold resistance ratio at {TEX}$LC_{50}${/TEX} to S strain. The resistance was started to in incline slowly to the resistance level of S strain after one year, and the resistance ratio was 4-fold in the 7 years after then. PCR-amplification of T. urticae DNA showed polymorphism in the amplifications with 12 primers in 100 kinds of arbitrary DNA sequences. RAPD amplification with primer OPR-12 (5`-ACAGGTGCGT-3`) showed amplified bands at 1,000 base pair in the S-and RS-strain, and at 350 base pair in R-strain. The results of polymorphism are genetic variabilities derived from development and selection of resistance in each strain. The peculiarly amplified fragments were guessed to participate in dicofol resistance. From the analysis of genetic similarity, it is inferred the gene composition of S-and RS-strain is much closer than that of R-strain.

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Characterization and cDNA Cloning of a Defensin-Like Peptide, Harmoniasin, from Harmonia axyridis

  • Kim, In-Woo;Lee, Joon-Ha;Park, Ha-Yan;Kwon, Young-Nam;Yun, Eun-Young;Nam, Sung-Hee;Kim, Seong-Ryul;Ahn, Mi-Young;Hwang, Jae Sam
    • Journal of Microbiology and Biotechnology
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    • v.22 no.11
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    • pp.1588-1590
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    • 2012
  • We compared the mRNA expression profile of the Harmonia axyridis larvae that were either untreated or treated with LPS. The extracted mRNAs were subjected to ACP RT-PCR analysis using a combination of arbitrary primers and oligo (dT) primer. Among the 47 DEGs differentially expressed, we identified a cDNA showing homology with defensin-like antibacterial peptide. The cDNA showed a putative 32-residue signal sequence and a 50-residue mature peptide named harmoniasin. We also investigated the antibacterial activity of the harmoniasin analog, which exhibited potent antibacterial activities against Gramnegative and -positive bacteria strains and it also evidenced no hemolytic activity.

A Genetic Marker for the Korean Native Cattle (Hanwoo) Found by an Arbitrarily Primed-Polymerase Chain Reaction (AP-PCR)

  • Lee, Ji-Seon;Lee, Chang-Hee;Nam, Doo-Hyun;Jung, Young-Ja;Yeo, Jung-Sou
    • BMB Reports
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    • v.33 no.3
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    • pp.208-212
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    • 2000
  • In order to develop a specific genetic marker for the Korean native cattle (Hanwoo), an arbitrarily-primed polymerase chain reaction (AP-PCR) analysis of 6 different cattle breeds was attempted. Eight different arbitrary primers, each longer than 20-mer nucleotides, were used. In comparison to the AP-PCR patterns, several distinctive DNA bands that are specific for a certain breed were detected. When the primer Kpn-X was employed, a 280bp DNA fragment was found to be specific only for Hanwoo. In an individual analysis of Hanwoo, this AP-PCR marker was observed in 123 head of cattle among the 153 that were tested (80.4%). Nucleotide sequencing revealed that this fragment has a short microsatellite sequence of tandem repeat, $A(G)_{1-2}\;(C)_{1-3}AGAG$. According to the analysis of AP-PCR band patterns, Hanwoo was discovered to be genetically most closely-related with Holstein among the various cattle breeds.

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Genetic Variability Based on Randomly Amplified Polymorphic DNA in Mistletoe Fig (Ficus deltoidea Jack) Collected from Peninsular Malaysia

  • Bhore, Subhash Janardhan;Arneida H., Nurul;Shah, Farida Habib
    • Journal of Forest and Environmental Science
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    • v.25 no.1
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    • pp.57-65
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    • 2009
  • Ficus deltoidea Jack is an important and popular medicinal plant species found in the Malaysia. Plants are being collected and used based on morphology and authentication to prevent adulteration is not in practice. In this study, twenty-six accessions of F. deltoidea Jack were collected from Kelantan and Terengganu states of Peninsular Malaysia to examine their genetic similarities and differences using randomly amplified polymorphic DNA (RAPD) technique. Out of 20 arbitrary primers, two primers (D-10 and D-11) were selected which produced reliable DNA polymorphism. D-10 and D-11 primers generated 138 RAPD bands ranging from 250 bp to 3000 bp. Ninety-nine of them were polymorphic loci (72%) and thirty-nine were nonpolymorphic loci (28%). A total of 56 bands with polymorphic loci were amplified with primer D-10 and analyzed by cluster analysis and UPGMA to present a dendrogram depicting the degree of genetic relationship among 26 accessions. Eight RAPD markers were sequenced to determine their identity. RAPD analysis showed the genetic diversity among 26 accessions of F. deltoidea Jack. The RAPD profile and RAPD marker sequences reported in this paper could be used in plant and/or plant material authentication. This study also suggested that RAPD can be a useful technique to study DNA polymorphism in F. deltoidea Jack.

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