National Institute of Agrobiological Sciences

Nitto Boseki Co., Ltd.

National Agriculture and Food Research Organization

Hideki Sezutsu

Toshiki Tamura

Isao Kobayashi

Keiro Uchino

Toshihide Miura

Iwao Kiyokawa

Ryo Kojima

Takuya Tsubota

Yuji Arimatsu

Katsuhiro Katayama

Taiyo Yoshioka

Tsunenori Kameda

Takao Suzuki

Tatsuya Ohashi

Sachiko Machida

Toshio Kanda

Katsura Kojima

Hiromitsu Tanaka

Kenichi Nakajima

Tetsuya Iizuka

Naoyuki Yonemura

Ken-ichiro Tatematsu

Toshiro Kobori

Mari Yamamoto

Mayumi Kameyama

Miyuki Kuramochi

The present invention pertains to a silkworm-type biotinylated CTLD14 or sRAGE and a method for manufacturing the same. One embodiment of the present invention provides a method for manufacturing biotinylated proteins, wherein the method includes A) a step for inserting a nucleic acid molecule for coding biotin ligase and protein in a coexpressable manner into a silkworm or a living organism that imparts sugar chains that are the same as the sugar chains of the silkworm, B) a step for causing the biotin ligase and protein to be expressed by disposing the silkworm or the living organism that imparts sugar chains that are the same as the sugar chains of the silkworm to conditions with which the nucleic acid molecule will carry out expression, and C) a step for administering biotin to the living organism and obtaining the biotinylated protein.

Biotinylated and oxidized LDL receptor and advanced glycation end product receptor produced using genetically engineered silkworm

A method of producing a transgenic silkworm that spins bagworm silks and producing a large quantity of bagworm silks by transgenic technology is developed and provided. A gene encoding a modified bagworm Fib H and a transgenic silkworm in which the gene is introduced, wherein the gene is obtained by cloning a gene fragment encoding a bagworm Fib H-like polypeptide comprising a partial amino acid sequence of bagworm Fib H, and fusing the gene fragment to a gene fragment encoding silkworm-derived Fib H, are provided.

Recombinant bagworm silk

This invention is intended to develop a promoter that can strongly induce marker gene expression throughout an embryo, so as to simply, efficiently, and accurately identify a transgenic insect at an early developmental stage, and to provide a gene expression vector into which such promoter has been incorporated as a transformant discrimination marker. Such exogenous gene expression vector comprises a polynucleotide comprising the nucleotide sequence as shown in SEQ ID NO: 1 as a promoter.

Exogenous gene expression vector, transformant discrimination marker, and transformant

Pentameric CRP is produced at a high efficiency by transferring DNA, which encodes monomeric CRP, into a silkworm to thereby construct a transgenic silkworm and then collecting and purifying pentameric CRP that is produced by the transgenic silkworm constructed above.

DNA encoding canine monomeric CRP and expression vector containing the DNA

Pentameric CRP-producing transgenic silkworm

Silkworms which have (i) a DNA encoding a transcriptional regulator operably linked downstream of a promoter of a DNA encoding a protein specifically expressed in the silk gland and (ii) a DNA encoding TRACP5 operably linked downstream of a target promoter of the transcriptional regulator were produced. The result showed that active TRACP5b was produced from the silkworms. This means that TRACP5 produced from the silk gland of the silkworms undergoes processing in the silk gland that is similar to the processing taking place at bone resorption sites.

Method for production of TRACP5b

Transgenic silkworms comprising GFP whose expression is regulated by the sericin gene promoter were produced. Observation of the silk glands of the last instar larvae of the silkworms showed fluorescence only in the middle silk glands. GFP was secreted from middle silkgland cells from around the spinning stage, indicating that GFP moved into the gland lumen. Finally, GFP was spun into cocoon filaments, and cocoons containing large amounts of GFP were produced. Thus, by using the promoter region of the sericin gene, recombinant proteins can be produced in the middle silk glands. Furthermore, the recombinant proteins produced in the middle silk glands were readily secreted into the lumen of the middle silk glands.

Methods for producing proteins using silkworm middle silk gland-specific gene expression system

Growing community of inventors

Tsukuba, Japan

Hideki Sezutsu