Expression of human growth hormone by the eukaryotic alga, Chlorella.
A method to use Chlorella to express a recombinant heterologous protein that can be recovered from the extracellular medium has been developed. Plasmids are constructed with an extracellular secretion signal sequence inserted between a promoter region and a gene for human growth hormone (hGH). The plasmids also contain a Kanr region which confers resistance to the antibiotic G418. Protoplasts are prepared by enzymatic treatment, and the plasmid is introduced by incubation of the protoplasts with polyethylene glycol and dimethyl sulfoxide. Cells are then grown in the presence of G418, and the medium is collected from 6 days after transfection. hGH is measured by immunoassay, and values for expressed hGH of about 200-600 ng/ml are obtained.
Hawkins
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Future Project Division, Toyota Motor Corporation, 1, Toyota-cho, Toyota, Aichi 471-8572, Japan.
Nakamura
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D000595:Amino Acid Sequence; D000900:Anti-Bacterial Agents / Q000494:pharmacology; D015139:Blotting, Southern; D015153:Blotting, Western; D002708:Chlorella / Q000187:drug effects / Q000235:genetics* / Q000378:metabolism; D002738:Chloroquine / Q000494:pharmacology; D003360:Cosmids; D018274:Electroporation; D015870:Gene Expression; D005839:Gentamicins / Q000494:pharmacology; D006358:Hot Temperature; D019382:Human Growth Hormone / Q000096:biosynthesis / Q000235:genetics*; D007438:Introns; D008969:Molecular Sequence Data; D010957:Plasmids; D011401:Promoter Regions, Genetic; D021382:Protein Sorting Signals; D014162:Transfection
D000900:Anti-Bacterial Agents; D005839:Gentamicins; D021382:Protein Sorting Signals; D019382:Human Growth Hormone; D002738:Chloroquine; C010680:antibiotic G 418