Production of an active feline interferon in the cocoon of transgenic silkworms using the fibroin H-chain expression system.
We constructed the fibroin H-chain expression system to produce recombinant proteins in the cocoon of transgenic silkworms. Feline interferon (FeIFN) was used for production and to assess the quality of the product. Two types of FeIFN fusion protein, each with N- and C-terminal sequences of the fibroin H-chain, were designed to be secreted into the lumen of the posterior silk glands. The expression of the FeIFN/H-chain fusion gene was regulated by the fibroin H-chain promoter domain. The transgenic silkworms introduced these constructs with the piggyBac transposon-derived vector, which produced the normal sized cocoons containing each FeIFN/H-chain fusion protein. Although the native-protein produced by transgenic silkworms have almost no antiviral activity, the proteins after the treatment with PreScission protease to eliminate fibroin H-chain derived N- and C-terminal sequences from the products, had very high antiviral activity. This H-chain expression system, using transgenic silkworms, could be an alternative method to produce an active recombinant protein and silk-based biomaterials.
Kurihara
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H
Toray Industries, Inc., New Frontiers Research Laboratories, 1111 Tebiro, Kamakura, Kanagawa 248-8555, Japan. Hiroyuki_Kurihara@nts.toray.co.jp
Sezutsu
H
H
Tamura
T
T
Yamada
K
K
D000595:Amino Acid Sequence; D000818:Animals; D030801:Animals, Genetically Modified; D012831:Bombyx / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D002415:Cats; D005349:Fibroins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D015870:Gene Expression; D007372:Interferons / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics; D008969:Molecular Sequence Data; D011401:Promoter Regions, Genetic / Q000235:genetics; D011679:Pupa / Q000235:genetics / Q000378:metabolism; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism
D011994:Recombinant Proteins; D005349:Fibroins; D007372:Interferons