Journal of Life Science (생명과학회지)

Korean Society of Life Science (한국생명과학회)
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Construction and Analysis of cDNA Library from Porphyra yezoensis

방사무늬 김의 cDNA Library 제조 및 분석

  • 서수분 (부산대학교 자연과학대학 분자생물학과) ;
  • 이은경 (부산대학교 자연과학대학 분자생물학과) ;
  • 김영진 (부산대학교 자연과학대학 분자생물학과)
  • Published : 1999.10.01
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Abstract

As an attempt to preserve resources in marine biological organisms, we first constructed a cDNA library from the red alga Porphyra yezoensis. The library construction method from P. yezoensis consists of three steps; those include protoplast presparation, RNA isolation, and phage library construction. Protoplast was prepared in order to remove much of the carbohydrate compounds which are characteristics of algal cell walls. Carbohydrate contamination in the purified RNA may inhibit further enzyme reactions, those carbohydrates should be removed. RNA samples prepared from protoplast still seemed to contain residual amount of carbohydrate because mRNA isolation with conventional method failed. We therefore developed a method with Poly ATtract mRNA isolation system. The constructed phage library was tested by analyzing cDNA insert in phage vector from randomly picked ten independent white plagues. All of the phages contained cDNA inserts with sizes ranging 0.5kb and 2.0kb.

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References

  1. J. Phycol. v.7 Secretion of vitamins and amino acids into the enviroment by Ochromononas danica Araonson,S. ;B. deAngelie ;O.Frank;H. Baker
  2. Anal. Biochem. v.154 Isolation of DNA from fungi mycelia and sclerotia without use of density gradient ultracentrifugation Biel. S.W.;F.W. Parrish
  3. J. Phycol. v.24 The use of plasmid DNA restriction endonuclease elineating redalgal species and population Goff. L.J.;A.W. Coleman
  4. Curr. Genet. v.18 Red algal plamids Goff,L.J.;A.W. Coleman
  5. Anal. Biochem. v.114 Lithium choride extraction of DNA from the seaweed Prophyra perforta(Rhodophyta) Holmes, D.S. Quigley
  6. J. Phycol. v.28 Lithium chlorice extraction of DNA from the seaweed Porphyra pertorta(Rhodophyta) Hong, Y.;D.A. Coury;M. Polne-Fuller and A. Gibor
  7. Anal. Biolchem. v.195 Isolation of nucleic acids from plants by different solvent prepartion Manning K.
  8. Planta(Berl.) v.14 Preparation of protoplasts from the green alga Enteromopha intestinalis Miller, P.;M.Callow;L. Evans
  9. Plant. Sci. v.104 Characterization of catalase from the seaweed Porphyra yezoensis Nakano. T.;M. Watanabe;M. Sato
  10. Plant. Physiol. v.60 Factors affecting extraction of intact ribonucleic acid from plant tissues containing interfering phenolic compounds Newbury,H.J.;V.J. Possingham
  11. J. Phycol. v.20 Developmental studies in Porphyra.I. Blade differentiation in Porphyra perforata as expressed by morphology . enzymatic digestion and protoplast regeneration Polne-Fuller, M.;A, Gibor