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KMID : 0545119910010040232
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Journal of Microbiology and Biotechnology
1991 Volume.1 No. 4 p.232 ~ p.239
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Purification and In Vitro Translation of Penicillium verruculosum Cellulase mRNA
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KIM, JEONG HO
CHUNG, KI CHUL/KANG, HYEN SAM/LEE, YOUNG KYU
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Abstract
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Caboxymethyl cellulase (CMCase) I was purified from the induced culture filtrate of Penicllium verruculosum F-3 by ammonium sulfate precipitation, DEAE-Sephadex A-50 chromatography and Bio-gel P-150 filtration. The purified enzyme was assumed to be a glycoprotein consisting of 8.5% carbohydrate and having a molecular weight of 70.000 in SDS-polycrylamide gel electrophoresis (SDS-PAGE). The purified enzyme-specific anti-CMCase I IgG was obtained by rabbit immunization and protein A-sepharose CL-4B chromatography. The fungal poly(A^+) RNA was isolated from the total RNA of the mycelium grown under cellulase induction conditions by oligo(dT)-cellulosse chromatography. The translation products in vitro were prepared by translating the isolated poly(A^+) RNA in rabbit reticulocyte lysate and analyzed by SDS-PAGE and fluorography. Of the translation products, CMCase I was identified by the immunoprecipitation against antiCMCase I IgG.
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