High Level Secretion bySaccharomyces cerevisiaeof Human Apolipoprotein E as a Fusion toRhizomucorRennin
| Autor: | Hisashi Yamada, Nobuhiko Nomura, Sueharu Horinouchi, Nobuyuki Matsubara, Teruhiko Beppu |
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| Rok vydání: | 1995 |
| Předmět: |
Blotting
Western Genetic Vectors Molecular Sequence Data Saccharomyces cerevisiae Biology Applied Microbiology and Biotechnology Biochemistry Analytical Chemistry law.invention Fusion gene Apolipoproteins E law Complementary DNA Gene expression Humans Amino Acid Sequence Cloning Molecular Molecular Biology Gene Base Sequence Organic Chemistry Nucleic acid sequence General Medicine biology.organism_classification Molecular biology Recombinant Proteins Yeast Blotting Southern Mucorales Recombinant DNA Chymosin Plasmids Biotechnology |
| Zdroj: | Bioscience, Biotechnology, and Biochemistry. 59:382-387 |
| ISSN: | 1347-6947 0916-8451 |
| DOI: | 10.1271/bbb.59.382 |
| Popis: | As the first step for production of human apolipoprotein E (hApoE) in Saccharomyces cerevisiae, the hApoE cDNA was cloned in Escherichia coli, on the basis of the nucleotide sequence reported previously. When the hApoE cDNA including its pre-sequence-encoding region was expressed under the control of the GAL7 promoter, no protein immunoreactive with anti-hApoE antibody was detected either in the culture medium or inside the cells. For efficient production and secretion of hApoE in S. cerevisiae, the mature hApoE-encoding region was fused to the prepro-sequence region of Rhizomucor rennin (MPR) and to the whole MPR gene including its prepro- and mature-MPR regions. When the fusion gene consisting of the prepro-sequence-encoding region and hApoE regions was expressed in S. cerevisiae, no protein reactive with the anti-hApoE antibody was detected in any fraction of the yeast cells, probably due to rapid degradation of the hApoE protein by yeast proteases. On the other hand, when hApoE was expressed as a fusion to the whole MPR protein, a considerable amount of the fused protein was secreted into the medium. The prepro-sequence of MPR was correctly processed from the fused protein in the medium by autocatalytic activity of MPR and by a protease(s) of the host cell.(ABSTRACT TRUNCATED AT 250 WORDS) |
| Databáze: | OpenAIRE |
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