Primary structure of the silk fibroin light chain determined by cDNA sequencing and peptide analysis

Yamaguchi, Kazunori; Kikuchi, Yoshimi; Takagi, Takashi; Kikuchi, Azusa; Oyama, Fumitaka; Shimura, Kensuke; Mizuno, Shigeki

doi:10.1016/0022-2836(89)90295-7

Cited by 175 publications

(127 citation statements)

References 35 publications

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“…지금까지 보고된 실크단백질의 구성은 피브 로인이 약 75%이고 세리신이 나머지 25%이고, 그 중에 서 피브로인은 heavy chain(H-chain), light-chain(L-chain) 및 fibrohexamerin 3개의 subunit 복합체가 6 : 6 : 1의 분자 비율로 구성되어 있다 (Inoue et al 2005, Yamaguchi et al 1989). 피브로인 단백질의 기본 구성은 H-chain 과 L-chain 이 1 : 1 비율로서 이황화결합을 이루고 있으며, 결합된 H-L chain 여섯 분자는 한 분자의 P25에 의해 비공유성 결 합으로 조립되어 있다 (Inoue et al 2005, Tanaka et al 1999 (Yamada et al 2001, Zhao et al 2009, Zhou et al 2003.…”

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Construction of fluorescent red silk using fibroin H-chain expression system

Kim¹,

Yun²,

Choi³

et al. 2012

Korean Journal of Sericultural Science

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We constructed the fibroin H-chain expression system to produce Discosoma sp. red fluorescent protein variant2 (DsRed2) in transgenic silkworm cocoon. Fluorescent cocoon could be made by fusing DsRed2 cDNA to the heavy chain gene and injecting it into a silkworm. The DsRed2 fusion protein, each with N-and C-terminal sequences of the fibroin H-chain, was designed to be secreted into the lumen of the posterior silk glands. The expression of the DsRed2/ H-chain fusion gene was regulated by the fibroin H-chain promoter. The use of the 3xP3-driven EGFP cDNA as a marker allowed us to rapidly distinguish transgenic silkworms. The EGFP fluorescence became visible in the ocelli and in the central and peripheral nervous system on the seventh day of embryonic development. A mixture of the donor and helper vector was micro-injected into 1,020 Kumokjam, bivoltin silkworm eggs. We obtained 6 broods. The cocoon was displayed strong red fluorescence, proving that the fusion protein was present in the cocoon. Accordingly, we suggest that the DsRed2 fluorescence silk will enable the production of novel biomaterial based on the transgenic silk.

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Construction of fluorescent red silk using fibroin H-chain expression system

Kim¹,

Yun²,

Choi³

et al. 2012

Korean Journal of Sericultural Science

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“…mori silk fibroin comprises BmFib-H, BmFib-L, and BmP25 proteins (Sprague, 1975;Couble et al, 1983;Yamaguchi et al, 1989), which are produced abundantly in the silk glands of 5th instar larvae . However, in this experiment bmo-miR-0001 and bmo-miR-0015 downregulated the expression of the BmFib-L gene.…”

Section: Discussionmentioning

confidence: 99%

“…The ASG is a catheter of silk protein with no function in silk protein biosynthesis and the MSG mainly synthesizes and secretes sericin proteins, whereas the PSG synthesizes and secretes silk fibroin, which accounts for more than 70% of the total silk protein. Fibroin comprises the fibroin heavy chain protein (Fib-H), fibroin light chain protein (Fib-L), and P25/fibrohexamerin protein (P25) (Sprague, 1975;Yamaguchi et al, 1989). Studies have shown that B. mori miRNAs are involved in regulation of fibroin gene expression.…”

Section: Introductionmentioning

confidence: 99%

bmo-miR-0001 and bmo-miR-0015 down-regulate expression of Bombyx mori fibroin light chain gene in vitro

Chen

Fan

Wang

et al. 2016

J. Zhejiang Univ. Sci. B

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Abstract:Based on bioinformatic analysis, we selected two novel microRNAs (miRNAs), bmo-miR-0001 and bmomiR-0015, from high-throughput sequencing of the Bombyx mori larval posterior silk gland (PSG). Firstly, we examined the expression of bmo-miR-0001 and bmo-miR-0015 in 12 different tissues of the 5th instar Day-3 larvae of the silkworm. The results showed that the expression levels of both bmo-miR-0001 and bmo-miR-0015 were obviously higher in the PSG than in other tissues, implying there is a spatio-temporal condition for bmo-miR-0001 and bmo-miR-0015 to regulate the expression of BmFib-L. To test this hypothesis, we constructed pri-bmo-miR-0001 expressing the plasmid pcDNA3.0 [ie1-egfp-pri-bmo-miR-0001-SV40] and pri-bmo-miR-0015 expressing the plasmid pcDNA3.0 [ie1-egfp-pribmo-miR-0015-SV40]. Finally, the BmN cells were harvested and luciferase activity was detected. The results showed that luciferase activity was reduced significantly (P<0.05) in BmN cells co-transfected by pcDNA3.0 [ie1-egfp-pri-bmomiR-0001-SV40] or pcDNA3.0 [ie1-egfp-pri-bmo-miR-0015-SV40] with pGL3.0 [A3-luc-Fib-L-3'UTR-SV40], suggesting that both bmo-miR-0001 and bmo-miR-0015 can down-regulate the expression of BmFib-L in vitro.

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“…Research on silkworm larval development and its applications is important for silk production and could improve the amount and quality of silk thread produced commercially (Li et al, 2005). Silk fibroin is secreted into the lumen of the posterior silk gland of the B. mori silkworm and is composed mainly of three polypeptides: a 350-kDa heavy chain (Ohmachi et al, 1982), a 26-kDa light chain (Yamaguchi et al, 1989;Kikuchi et al, 1992) and fibrohexamerin (Inoue et al, 2000). The fibroin light chain gene, fib-L, is located on chromosome 14 and consists of 14,626 bp, containing seven exons with large-sized introns (Kikuchi et al, 1992).…”

Section: Introductionmentioning

confidence: 99%

Allelic variability in the third intron of the fibroin light chain gene in Bombyx mori (Lepidoptera: Bombycidae)

Barbosa¹,

Bravo²,

Zanatta³

et al. 2009

Genet. Mol. Res.

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ABSTRACT. Conformation-sensitive gel electrophoresis is a useful method for identifying allele polymorphism; it provides co-dominant molecular markers. Using this method, we identified genetic variability in the third intron of the fibroin light chain gene, fib-L, in six Bombyx mori strains. Only Chinese C21A strain did not demonstrate allelic alterations, showing only homoduplex DNA molecules. We found distinct heteroduplex profiles in the Japanese HAA, M12B and M19-2 and the Chinese C25B and C24-2 strains. Analysis with restriction endonuclease fingerprinting conformation-sensitive gel electrophoresis demonstrated the potential of this method for the identification of allelic variability in B. mori; this was confirmed by cloning and sequencing the different alleles. The main alteration was a 12-bp deletion in two alleles of the C24-2 strain and one allele of the HAA strain; this deletion results in specific heteroduplex DNA molecule profiles.

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Primary structure of the silk fibroin light chain determined by cDNA sequencing and peptide analysis

Cited by 175 publications

References 35 publications

Construction of fluorescent red silk using fibroin H-chain expression system

Construction of fluorescent red silk using fibroin H-chain expression system

bmo-miR-0001 and bmo-miR-0015 down-regulate expression of Bombyx mori fibroin light chain gene in vitro

Allelic variability in the third intron of the fibroin light chain gene in Bombyx mori (Lepidoptera: Bombycidae)

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