• Title/Summary/Keyword: Species-specific PCR

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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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Rapid Detection and Discrimination of the Three Salmonella Serotypes, S. Pullorum, S. Gallinarum and S. Enteritidis by PCR-RFLP of ITS and fliC Genes

  • Cha, Se-Yeoun;Jang, Du-Hee;Kim, Sang-Min;Park, Jong-Beom;Jang, Hyung-Kwan
    • Korean Journal of Poultry Science
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    • v.35 no.1
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    • pp.9-13
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    • 2008
  • Salmonella enterica serotype gallinarum biovar Gallinarum or Pullorum and Salmonella enterica serotype Enteritidis are the most important diseases in poultry industry. Transitional diagnosis methods of these diseases such as direct isolation and identification by a biochemical test are time consuming with low specificity. In this study, we have focused on the suitable procedure for the rapid and accurate diagnosis of diseases derived from the three Salmonella strains. We initially confirmed Salmonella species by PCR using a specific ITSF/ITSR primer pair instead of biochemical test, and then the PCR-amplified phase 1 flagellin (fliC) using a specific fliCF/fliCR primer pair was digested with a restriction endonuclease, Bpm I and/or Bfa I, to discriminate among S. Pullorum, S. Gallinarum, and S. Enteritidis. We found that these methods could be applied to field isolates of the three Salmonella strains to detect and to discriminate rapidly for convenient diagnosis.

RAPD-SCAR Markers Linked to Medium-Leaf Zoysiagrass Ecotypes (한국잔디 중지 변이개체와 연관된 RAPD-SCAR 마커)

  • Chung, Sung Jin;Park, Su Jeong;Kim, Hun Joong;Yang, Geun-Mo;Choi, Joon-Soo;Oh, Chan-Jin;Jang, Deok-Hwan;Song, In-Ja;Lee, Geung-Joo
    • Weed & Turfgrass Science
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    • v.2 no.2
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    • pp.191-197
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    • 2013
  • Two medium-leaf ecotypes (CY6069, CY6097) belonging to one species (Zoysia japonica) of Korean lawngrasses were selected in sod production fields in Jang Seong, Korea. They were reported to have distinct morphological and growth rate characteristics different from the preferred medium-leaf type zoysiagrass in Korea. This study was conducted to define further the genotypic difference at the molecular level and to develop DNA marker based on the specific DNA fragment. Polymorphic DNA fragments were first explored by using randomly amplified polymorphic DNA (RAPD) primers, which were then converted into PCR-based sequence characterized amplified region (SCAR) markers. The CY6069-specific primer set amplified about 550 bp successfully, while the CY6097 marker produced the expected 690 bp band, by which those markers were nominated by CY6069_550 and CY6069_690 SCARs, respectively. Together with the reported morphological and other phenotypic features, the SCAR markers confirmed in this study will be useful to identify those medium-leaf zoysiagrass genotypes when they are cultivated with other vegetatively propagated warm-season turfgrasses in sod farms.

Pathogenicity and PCR detection of Vibrio tapetis in Manila clams, Ruditapes philippinarum (양식 바지락, Ruditapes philippinarum에 대한 Vibrio tapetis의 병원성과 PCR법에 의한 진단)

  • Park, Sung-Woo;Lee, Kyung-Hee
    • Journal of fish pathology
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    • v.18 no.1
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    • pp.39-48
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    • 2005
  • Pathogenicity of Vibrio tapetis, the causative bacterium of 'brown ring disease (BRD)' was evaluated in Manila clams (Ruditapes philippinarumi by artificially 0.1 $m\ell$ infection of $1.0\times10^5$cells and $1.0\times10^8$ cells at 20 $^{\circ}C$. A PCR assay based on 16S rRNA to detect the bacteria in clam tissues was established. Accumulative mortality of clams infected with $1.0\times10^7$cells and $1.0\times10^4$ cells per an individual of the bacteria was 67.5% and 7.5%, respectively. However, the deposit of brown pigment in the inner shells by accumulation of chonchiolin was not found. The bacteria were not be able to re-isolate from the infected clams by the conventional agar plate method but were easily detected by PCR assay established in this experiment. In clams artificially infected with 10 species of Vibrio, a 414bp for V. tapetis was detected in PCR assay. The specific band in the clams infected with $1.0\times10^4$cells per an individual of V. tapetis was detected only in gills one day after the infection but never be found in any tissues including gills three days after the infection. In the case of clams infected with $1.0\times10^8$cells per an individual of V. tapetis the specific band was detected in gills and intestine one day after the infection, in all tissues three days after the infection, and then in gills and adductor muscle nine days after the infection. The PCR assay was applied to detect V. tapetis in manila clam, surf clam (Mactra veneriformis), oyster (Crassostrea gigas) and Thomas' rapa whelk (Rapana venosa) taken from Taean and Gochang from April to July 2004. The infection rates were detected to 23.1% and 9.4% in the oyster and surf clam, while manila clam and Thomas' rapa whelk were not found.

Identification of Two Isoforms of Aminopeptidase N in Aedes aegypti Larval Midgut

  • Pootanakit, Kusol;Angsuthanasombat, Chanan;Panyim, Sakol
    • BMB Reports
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    • v.36 no.5
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    • pp.508-513
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    • 2003
  • The bacterium Bacillus thuringiensis produces toxin inclusions that are deleterious to target insect larvae. These toxins are believed to interact with a specific receptor protein(s) that is present on the gut epithelial cells of the larvae. In various insect species (in particular those belonging to the lepidopteran class), aminopeptidase N (APN) is one of the two receptor proteins that are considered to be involved in toxin-receptor interactions. However, in mosquitoes, the nature and identity of the receptor protein is unknown. Here, using RT-PCR, we identified two isoforms of the APN transcripts in the Aedes aegypti mosquito larval midgut. These results are congruent with a previous report of multiple isoforms of the APN gene expression in lepidopteran larvae. Which of the two isoforms (or other yet unidentified receptor proteins) is involved in the killing of mosquito larvae remains to be elucidated.

A Study of Tissue-Specific Expression and Analysis of Structure and DNA Sequences of Porcine Cytochrome P450 Aromatase Genes

  • Ghimeray, Amal-Kumar;Chae, Sung-Hwa;Lee, Eun-Ju;Quan Feng;Nina Oriero;Park, Inho
    • Proceedings of the KSAR Conference
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    • 2004.06a
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    • pp.196-196
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    • 2004
  • By use of RT-PCR coupled with DHPLC technique (WAVE analysis), pattern of isoforms of porcine cytochrome P450 aromatase gene was investigated. Relatively higher expression of aromatase mRNA was observed in testis than in ovary and this result accounted for the previous findings of higher blood estrogen level in male compared with female in this species. (omitted)

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Method Development for Electrotransformation of Acidithiobacillus caldus

  • Chen, Linxu;Lin, Jianqun;Li, Bing;Lin, Jianqiang;Liu, Xiangmei
    • Journal of Microbiology and Biotechnology
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    • v.20 no.1
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    • pp.39-44
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    • 2010
  • Acidithiobacillus caldus is an acidophilic, chemolithotrophic bacterium that plays an important role in bioleaching. Gene transformation into A. caldus is difficult, and only the conjugation method was reported successful, which was a relatively sophisticated method. In this research, electrotransformation of A. caldus species was achieved for the first time using A. caldus Y-3 and plasmid pJRD215. Transformants were confirmed by colony PCR specific to the str gene on pJRD215, and the recovery of the plasmid from the presumptive transformants. Optimizations were made and the transformation efficiency was increased from 0.8 to $3.6{\times}10^4$ transformants/${\mu}g$ plasmid DNA. The developed electrotransformation method was convenient in introducing foreign genes into A. caldus.

Cloning and expression of new laccase gene (soncotA) from Bacillus sonorensis KCTC13918 in E. coli (Bacillus sonorensis KCTC13918로부터 새로운 laccase유전자 (soncotA)의 클로닝과 대장균에서의 발현)

  • Choi, Shin-Geon;Yoon, Hyeonjong
    • Journal of Industrial Technology
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    • v.37 no.1
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    • pp.16-20
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    • 2017
  • A new putative laccase gene (soncotA) which show 78% homology with that from Bacillus licheniformis (liccotA) was isolated from draft genome sequence of Bacillus sonorensis KCTC 13918. A 1,545 bp of PCR product corresponding 514 amino acids was cloned into NdeI-NotI site of pET21c and expressed as soluble form in E. coli. About 59 kDa size of recombinant laccase was purified into homogenity by Ni-NTA column and laccase activity was confirmed by zymography. The enzymatic properties of recombinant laccase were characterized. The specific activity of B. sonorensis laccase was 0.033 fold lower than that of Bacillus licheniformis laccase. The finding of new laccase gene broadened the enzymatic diversity of Bacillus species laccases.

Prevalence of Torque teno viruses among pigs and cattle in Korea

  • Kim, Ji Yeon;Kweon, Chang Hee;Lee, Kyung Woo;Jeong, Wooseog;Jean, Young Hwa
    • Korean Journal of Veterinary Research
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    • v.50 no.1
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    • pp.55-57
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    • 2010
  • Torque teno virus (TTV), a species of Anellovirus, is a non-enveloped single stranded DNA virus with a wide range of animal hosts. The incidence of TTV is quite ubiquitous throughout the world. A total of 235 serum samples obtained from 137 pigs and 98 cattle at slaughterhouses in Korea during April 2005 to May 2005 were tested by TTV-specific PCR as to monitor prevalence of TTV among swine and cattle. As a result, the prevalent rates of TTVs in pigs and cattle were 43.1% and 4.1%, respectively. It seems that TTV infection is quite prevalent in swine population.

Systemic Infection of Maize, Sorghum, Rice, and Beet Seedlings with Fumonisin-Producing and Nonproducing Fusarium verticillioides Strains

  • Dastjerdi, Raana;Karlovsky, Petr
    • The Plant Pathology Journal
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    • v.31 no.4
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    • pp.334-342
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    • 2015
  • Two fumonisin-nonproducing strains of Fusarium verticillioides and their fumonisin producing progenitors were tested for aggressiveness toward maize, sorghum, rice, and beetroot seedlings grown under greenhouse conditions. None of the plants showed obvious disease symptoms after root dip inoculation. Fungal biomass was determined by species-specific real-time PCR. No significant (P = 0.05) differences in systemic colonization were detected between the wild type strains and mutants not producing fumonisins. F. verticillioides was not detected in any of the non-inoculated control plants. The fungus grew from roots to the first two internodes/leaves of maize, rice and beet regardless of fumonisin production. The systemic growth of F. verticillioides in sorghum was limited. The results showed that fumonisin production was not required for the infection of roots of maize, rice and beet by F. verticillioides.