New and highly efficient expression systems for expressing selectively foreign protein in the silk glands of transgenic silkworm

Zhao, Aichun; Zhao, Tianfu; Zhang, Yuansong; Xia, Qingyou; Lü, Cheng; Zhou, Zeyang; Zhang, Xiang; Nakagaki, Masayuki

doi:10.1007/s11248-009-9295-7

Cited by 60 publications

(73 citation statements)

References 32 publications

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“…Primers used here and hereafter are listed in Supplementary information, Table S1. The DNA vector (Fil-GAL4 or UAS-Ras1 CA ) and helper plasmid pHA3PIG were injected into silkworm eggs at a concentration of 0.4 μg/μl for each plasmid [31]. Positive G1 embryos were selected by DsRed fluorescence for Fil-GAL4 and EGFP fluorescence for UAS-Ras1 CA under an Olympus MVX10 fluorescence stereomicroscope [30,31].…”

Section: The Binary Gal4/uas Transgenic Silkworm Systemmentioning

confidence: 99%

“…The DNA vector (Fil-GAL4 or UAS-Ras1 CA ) and helper plasmid pHA3PIG were injected into silkworm eggs at a concentration of 0.4 μg/μl for each plasmid [31]. Positive G1 embryos were selected by DsRed fluorescence for Fil-GAL4 and EGFP fluorescence for UAS-Ras1 CA under an Olympus MVX10 fluorescence stereomicroscope [30,31]. To confirm the positive hits detected by fluorescence microscopy, the insertion sites of the transgenic Fil-GAL4 and UAS-Ras1 CA lines were also determined by inverse PCR [32].…”

Section: The Binary Gal4/uas Transgenic Silkworm Systemmentioning

confidence: 99%

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Ras1CA overexpression in the posterior silk gland improves silk yield

Zhu

et al. 2011

Cell Res

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Sericulture has been greatly advanced by applying hybrid breeding techniques to the domesticated silkworm, Bombyx mori, but has reached a plateau during the last decades. For the first time, we report improved silk yield in a GAL4/UAS transgenic silkworm. Overexpression of the Ras1 CA oncogene specifically in the posterior silk gland improved fibroin production and silk yield by 60%, while increasing food consumption by only 20%. Ras activation by Ras1 CA overexpression in the posterior silk gland enhanced phosphorylation levels of Ras downstream effector proteins, up-regulated fibroin mRNA levels, increased total DNA content, and stimulated endoreplication. Moreover, Ras1 activation increased cell and nuclei sizes, enriched subcellular organelles related to protein synthesis, and stimulated ribosome biogenesis for mRNA translation. We conclude that Ras1 activation increases cell size and protein synthesis in the posterior silk gland, leading to silk yield improvement.

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Section: The Binary Gal4/uas Transgenic Silkworm Systemmentioning

confidence: 99%

Section: The Binary Gal4/uas Transgenic Silkworm Systemmentioning

confidence: 99%

Ras1CA overexpression in the posterior silk gland improves silk yield

Zhu

et al. 2011

Cell Res

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“…Thus, the synthesized fusion protein might be integrated into the fibroin complex through an association with endogenous fibroin H-chains and the fibroin L-chain regions of the fusion protein, allowing it to be efficiently secreted from the cells into the cocoon. After this success, several improvements were reported for the promoter and fusion partner to efficiently express recombinant proteins in the PSG (Kojima et al 2007;Shimizu et al 2007;Zhao et al 2010). These studies offered experimental evidence for the production of cocoons containing recombinant proteins using the transgenic silkworm.…”

Section: Expression Of Recombinant Proteins In the Psgmentioning

confidence: 99%

Transgenic silkworms that weave recombinant proteins into silk cocoons

Tomita

2010

Biotechnol Lett

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As a result of breeding for more than 4,000 years, the silkworm, Bombyx mori, has acquired the ability to synthesize bulk amounts of silk proteins in its silk glands. To utilize this capacity for mass production of useful proteins, transgenic silkworms were generated that synthesized recombinant proteins in the silk gland and secreted them into the silk cocoon. The silk gland is classified into two main regions: the posterior (PSG) and the middle silk gland (MSG). By controlling the expressed regions of the recombinant protein gene in the silk gland, we were able to control the localization of the synthesized protein in the silk thread. Expression in the PSG or MSG led to localization in the insoluble fibroin core or hydrophilic outer sericin layer, respectively. This review focuses on the expression of recombinant protein in the MSG of transgenic silkworms. The recombinant protein secreted in the sericin layer is extractable from the cocoon with only a small amount of endogenous silk protein contamination by soaking the cocoon in mild aqueous solutions. The possibility of utilizing transgenic silkworms as a valuable tool for the mass production of therapeutic and industrially relevant recombinant proteins is discussed.

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“…An alternative way has been recently developed to avoid these shortcomings, where the genes of the Bombyx mori silkworm were screened and designed to directly obtain colorful silk from the sericterium of the silkworm [14]. Most recently, another "green" approach for the direct production of intrinsically colored and luminescent silk was also demonstrated by feeding domesticated silkworms organic dyes [15,16].…”

Section: Introductionmentioning

confidence: 99%