A New Method for the Modification of Fibroin Heavy Chain Protein in the Transgenic Silkworm

  title={A New Method for the Modification of Fibroin Heavy Chain Protein in the Transgenic Silkworm},
  author={Katsura Kojima and Yoshihiko Kuwana and Hideki Sezutsu and Isao Kobayashi and Keiro Uchino and Toshiki Tamura and Yasushi Tamada},
  journal={Bioscience, Biotechnology, and Biochemistry},
  pages={2943 - 2951}
We constructed a new plasmid vector for the production of a modified silk fibroin heavy chain protein (H-chain) in the transgenic silkworm. The plasmid (pHC-null) contained the promoter and the 3′ region of a gene encoding the H-chain and the coding regions for the N-terminal domain and the C-terminal domain of the H-chain. For the model protein, we cloned a foreign gene that encoded EGFP between the N-terminal domain and the C-terminal domain in pHC-null and generated transgenic silkworms that… 

New insight into the mechanism underlying fibroin secretion in silkworm, Bombyx mori

The results not only provide a theoretical basis for the genetic modification of silk fiber as a functional biomaterial but also are of great significance to establishing a new silk gland bioreactor to mass‐produce exogenous proteins in an active form.

Utilization of Transgenic Silkworms for Recombinant Protein Production

Recently developed methods for producing recombinant proteins in the Silkworm use the silk-synthesis systems in the silk gland and show that recombinant protein can be produced very efficiently.

Construction of transgenic silkworm spinning antibacterial silk with fluorescence

An investigation of the number of bacteria attached to a cocoon showed the transgenic silkworm cocoon possessed antibacterial properties, suggesting the performance of silk can be improved by modifying the fibroin gene.

Expression of the Japanese oak silkworm Antheraea yamamai fibroin gene in the domesticated silkworm Bombyx mori

Results indicate that the promoter function and splicing machinery are well conserved between A. yamamai and B. mori but that the secretion mechanism of fibroin is diversified between the two.

Transgenic silkworms that weave recombinant proteins into silk cocoons

  • M. Tomita
  • Chemistry, Medicine
    Biotechnology Letters
  • 2010
This review focuses on the expression of recombinant protein in the MSG of transgenic silkworms, which is a valuable tool for the mass production of therapeutic and industrially relevant recombinant proteins.

Construction of a binary transgenic gene expression system for recombinant protein production in the middle silk gland of the silkworm Bombyx mori

This binary system using a GAL4–Ser1 promoter construct is a useful tool for the mass production of recombinant proteins of biomedical and pharmaceutical interest in silkworm.

An efficient strategy for producing a stable, replaceable, highly efficient transgene expression system in silkworm, Bombyx mori

We developed an efficient strategy that combines a method for the post-integration elimination of all transposon sequences, a site-specific recombination system, and an optimized fibroin H-chain

Targeted glycoengineering extends the protein N-glycosylation pathway in the silkworm silk gland.

Production of scFv-Conjugated Affinity Silk Powder by Transgenic Silkworm Technology

The results strongly suggest the promise of scFv-conjugated silk fibroin as an alternative affinity reagent, which can be manufactured using transgenic silkworm technology at lower cost than traditional affinity carriers.

Production of scFv-conjugated affinity silk film and its application to a novel enzyme-linked immunosorbent assay

A transgenic silkworm that spins silk protein containing the fibroin L-chain linked with the single-chain variable fragment (scFv) as a fusion protein is generated that serves as the basis for an alternative immunodetection system.



Biosynthesis and cocoon-export of a recombinant globular protein in transgenic silkworms

A gene construct was made by fusing the coding sequence of the red fluorescent protein to the exon 2 of the fibrohexamerin gene, that encodes a subunit of fibroin, the major silk protein of the silkworm Bombyx mori, to establish a series of transgenic lines.

Transgenic silkworms produce recombinant human type III procollagen in cocoons

The viability of transgenic silkworms as a tool for producing useful proteins in bulk is demonstrated by immunoblotting, collagenase-sensitivity tests, and amino acid sequencing.

Production of a chimeric fibroin light-chain polypeptide in a fibroin secretion-deficient naked pupa mutant of the silkworm Bombyx mori.

The chimeric L-chain molecule of about 27 kDa is present in posterior silk glands of Nd-s and Nd -sD strains without disulfide-bonding to the fibroin heavy (H-) chain, as revealed by Western blotting with the antibody specific to the C-terminal half of the mutant L- chain.

Further evidence for importance of the subunit combination of silk fibroin in its efficient secretion from the posterior silk gland cells

These results, together with the previous results on the effect of the H chain gene-linked Nd(2) mutation, strongly suggest that the H-L subunit combination of silk fibroin is important for its efficient secretion.

Reduced level of secretion and absence of subunit combination for the fibroin synthesized by a mutant silkworm, Nd(2)

Evidence suggesting that the H chain derived from the Nd(2) allele is structurally abnormal is presented and how the H-L subunit structure is advantageous in the secretion of fibroin is discussed.

The sequence around the 5′ end of the fibroin gene from the wild silkworm, Bombyx mandarina, and comparison with that of the domesticated species, B. mori

The view that the B. mandarina which exists in nature at present is a possible ancestor of the domesticated silkworm, B. mori, is supported.