DC Field | Value | Language |
---|---|---|
dc.contributor.author | M J Kim | - |
dc.contributor.author | H J Chung | - |
dc.contributor.author | S M Park | - |
dc.contributor.author | Sung Goo Park | - |
dc.contributor.author | D K Chung | - |
dc.contributor.author | M S Yang | - |
dc.contributor.author | D H Kim | - |
dc.date.accessioned | 2017-04-19T09:01:17Z | - |
dc.date.available | 2017-04-19T09:01:17Z | - |
dc.date.issued | 2004 | - |
dc.identifier.issn | 1017-7825 | - |
dc.identifier.uri | https://oak.kribb.re.kr/handle/201005/6567 | - |
dc.description.abstract | On the foundation of a database of genome sequences and protein analyses, the ability to clone a gene based on a peptide analysis is becoming more feasible and effective for identifying a specific gene and its protein product of interest. As such, the current study conducted a protein analysis using 2-D PAGE followed by MALDI-TOF and ESI-MS to identify a highly expressed gene product of C. parasitica. A distinctive and highly expressed protein spot with a molecular size of 47.2 kDa was randomly selected and MALDI-TOF MS analysis was conducted. A homology search indicated that the protein appeared to be a fungal enolase (eno1). Meanwhile, multiple alignments of fungal enolases revealed a conserved amino acid sequence, from which degenerated primers were designed. A screening of the genomic λ library of C. parasitica, using the PCR amplicon as a probe, was conducted to obtain the full-length gene, while RT-PCR was performed for the cDNA. The E. coli-expressed eno1 exhibited enolase enzymatic activity, indicating that the cloned gene encoded the C. parasitica enolase. Moreover, ESI-MS of two of the separated peptides resolved from the protein spot on 2-D PAGE revealed sequences identical to the deduced sequences, suggesting that the cloned gene indeed encoded the resolved protein spot. Northern blot analysis indicated a consistent accumulation of an eno1 transcript during the cultivation. | - |
dc.publisher | Korea Soc-Assoc-Inst | - |
dc.title | Matrix-assisted laser desorption/ionization time-of-flight(MALDI-TOF)-based cloning of enolase, ENO1, from Cryphonectria parasitica | - |
dc.title.alternative | Matrix-assisted laser desorption/ionization time-of-flight(MALDI-TOF)-based cloning of enolase, ENO1, from Cryphonectria parasitica | - |
dc.type | Article | - |
dc.citation.title | Journal of Microbiology and Biotechnology | - |
dc.citation.number | 3 | - |
dc.citation.endPage | 627 | - |
dc.citation.startPage | 620 | - |
dc.citation.volume | 14 | - |
dc.contributor.affiliatedAuthor | Sung Goo Park | - |
dc.contributor.alternativeName | 김명주 | - |
dc.contributor.alternativeName | 정해종 | - |
dc.contributor.alternativeName | 박승문 | - |
dc.contributor.alternativeName | 박성구 | - |
dc.contributor.alternativeName | 정대균 | - |
dc.contributor.alternativeName | 양문식 | - |
dc.contributor.alternativeName | 김대혁 | - |
dc.identifier.bibliographicCitation | Journal of Microbiology and Biotechnology, vol. 14, no. 3, pp. 620-627 | - |
dc.subject.keyword | 2-D PAGE | - |
dc.subject.keyword | C. parasitica | - |
dc.subject.keyword | enolase | - |
dc.subject.keyword | ESI-MS | - |
dc.subject.keyword | MALDI-TOF | - |
dc.subject.local | 2-D PAGE | - |
dc.subject.local | 2D PAGE | - |
dc.subject.local | C. parasitica | - |
dc.subject.local | enolase | - |
dc.subject.local | ESI-MS | - |
dc.subject.local | MALDI-TOF | - |
dc.subject.local | MALDITOF | - |
dc.description.journalClass | Y | - |
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