KOPRI Repository

Cloning, expression and partial characterization of antifreeze protein from a psychrophilic yeast isolated from the Arctic

Cited 0 time in wos
Cited 0 time in scopus
Metadata Downloads

Full metadata record

DC FieldValueLanguage
dc.contributor.authorLee, Soo Young-
dc.contributor.authorPark, Kyoung Sun-
dc.contributor.authorKim, Hak Jun-
dc.contributor.authorKang, Sung-Ho-
dc.description.abstractA psychrophilic yeast having an extracellular antifreeze protein (AFP) was isolated from the Arctic. Its 18S and 28S ribosomal RNA sequences have a high similarity with Leucosporidium antarcticum but its ITS1, 5.8S, and ITS2 sequences are variable. We designated this isolate as L. sp. AY30. The semi-purified AFP from the culture medium of this yeast grown at 1 ℃ for 1 week has high antifreeze activity and is about 26 kDa in size on SDS-PAGE analysis. The genomic DNA sequence of AFP was obtained from a draft version of pyrosequencing data using local tBlastN. The candidate contig region of about 2 Kb in size containing the AFP gene was reamplified by PCR and sequenced. The forward upstream PCR primers for 3’-RACE were designed from the genomic DNA sequence. The largest 3’-RACE PCR product of about 1 Kb was sequenced. The nucleotide sequence contained 5’-UTR (119 bp) and 3’-UTR (40 bp). The ORF consists of 261 amino acids and the deduced protein was calculated as 26 kDa in size. The N-terminal 20 residues were predicted as a signal peptide. The mature AFP gene was inserted into pCold expression vector with N-terminal His-Tag and Factor Xa sequence. The recombinant plasmid was transformed into E. coli. The transformant was induced for 20 hours at 15℃ using 1 mM IPTG. The overexpressed AFP in the soluble faction of cell lysate was purified using Ni affinity chromatography. The ice crystal morphology and TH activity was analyzed using nanolitre osmometer. Its ice crystal shape and the burst shape revealed that it may bind to the basal plane of ice crystal, which is the characteristic of hyperactive AFPs. The TH activity was about 0.3 ℃ in the concentration of 3 ug/ml.-
dc.titleCloning, expression and partial characterization of antifreeze protein from a psychrophilic yeast isolated from the Arctic-
dc.title.alternative북극 유래 호냉성 효모로부터 결빙방지단백질 분석, 유전자 규명 및 발현에 관한 연구-
dc.identifier.bibliographicCitationLee, Soo Young, et al. 2009. Cloning, expression and partial characterization of antifreeze protein from a psychrophilic yeast isolated from the Arctic. 2009.07.19~.-
dc.subject.keywordantifreeze protein-
Appears in Collections  
2008-2010, Development of Longer preservation of Blood Using Antifreeze Molecules Derived from Polar Organisms (08-10) / Kim, Hak Jun
Files in This Item
There are no files associated with this item.
General Conditions
      ROMEO Green
    Can archive pre-print and post-print or publisher's version/PDF
      ROMEO Blue
    Can archive post-print (ie final draft post-refereeing) or publisher's version/PDF
      ROMEO Yellow
    Can archive pre-print (ie pre-refereeing)
      ROMEO White
    Archiving not formally supported

    Items in DSpace are protected by copyright, with all rights reserved, unless otherwise indicated.