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학술연구/단체지원/교육 등 연구자 활동을 지속하도록 DBpia가 지원하고 있어요.
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논문 기본 정보
- 자료유형
- 학술저널
- 저자정보
- 저널정보
- 한국미생물생명공학회 Journal of Microbiology and Biotechnology Journal of Microbiology and Biotechnology 제14권 제3호
- 발행연도
- 2004.1
- 수록면
- 620 - 627 (8page)
이용수
초록· 키워드
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On the foundation of a database of genomesequences and protein analyses, the ability to clone a genebased on a peptide analysis is becoming more feasible andeffective for identifying a specific gene and its proteinproduct of interest. As such, the current study conducted aprotein analysis using 2-D PAGE followed by MALDI-TOFand ESI-MS to identify a highly expressed gene product of C.parasitica. A distinctive and highly expressed protein spotwith a molecular size of 47.2 kDa was randomly selected andMALDI-TOF MS analysis was conducted. A homologysearch indicated that the protein appeared to be a fungalenolase (eno1). Meanwhile, multiple alignments of fungalenolases revealed a conserved amino acid sequence, fromwhich degenerated primers were designed. A screening of thegenomic l library of C. parasitica, using the PCR ampliconas a probe, was conducted to obtain the full-length gene,while RT-PCR was performed for the cDNA. The E. coliexpressedeno1 exhibited enolase enzymatic activity, indicatingthat the cloned gene encoded the C. parasitica enolase.Moreover, ESI-MS of two of the separated peptides resolvedfrom the protein spot on 2-D PAGE revealed sequencesidentical to the deduced sequences, suggesting that the clonedgene indeed encoded the resolved protein spot. Northern blotanalysis indicated a consistent accumulation of an eno1transcript during the cultivation.
목차
등록된 정보가 없습니다.
참고문헌 (27)
참고문헌 신청
Anagnostakis,S.L / 1982 / Biological control of chestnutblight 215 : 466 ~ 471
Brindle / 1990 / Multiple factors bind the upstreamactivation sites of the yeast enolase genes ENO1 and ENO2 binds cis-acting sequences which modulate repression oractivation
Brown / 1987 / The structure andexpression of nuclear gene in Saccharomyces cerevisiae Gene Structure in EukaryoticMicrobes : 1 ~ 26
Chen / 1996 / Fungalgene transcript accumulation and elevation of G-proteinregulatedcAMP levels by a virulence-attenuating hypovirus 93 : 7996 ~ 8000
Choi / 1992 / Molecularanalysis of the laccase gene from the chestnut blight fungusand selective suppression of its expression in an isogenichypovirulent strain : 119 ~ 128
Brindle / 1990 / Multiple factors bind the upstreamactivation sites of the yeast enolase genes ENO1 and ENO2 binds cis-acting sequences which modulate repression oractivation
Brown / 1987 / The structure andexpression of nuclear gene in Saccharomyces cerevisiae Gene Structure in EukaryoticMicrobes : 1 ~ 26
Chen / 1996 / Fungalgene transcript accumulation and elevation of G-proteinregulatedcAMP levels by a virulence-attenuating hypovirus 93 : 7996 ~ 8000
Choi / 1992 / Molecularanalysis of the laccase gene from the chestnut blight fungusand selective suppression of its expression in an isogenichypovirulent strain : 119 ~ 128
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