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

Insect Science 2009年第6期16卷 465-473页

作者： Isao Kobayashi Katsura Kojima Hideki Sezutsu keiro uchino Toshiki Tamura Transgenic Silkworm Research Center Silk Materials Research Unit National Institute of Agrobiological Sciences Ibaraki Japan

To understand the evolutionary conservation ofthe gene expression mechanism and secretion machinery between Antheraea and Bombyx fibroins, we introduced the genomic A. yamamai fibroin gene into the domesticated silkworm, B. mori. The spliced A. yamamai fibroin mRNA appeared only in the posterior region of the silk gland of the transgenic silkworm, suggesting that the functions of the fibroin promoter region and the splicing machinery are conserved between these two species. The A. yamarnai fibroin protein was detected in the lumen of the silk gland of the transgenic silkworm, albeit at lower levels compared with the B. mori-type fibroin. We found a strong degeneration of the posterior region of the silk gland of the transgenic silkworm. As a result, the cocoon shell weight was much lower in the transgenic silkworm than in the non-transgenic line. These 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.

关键词： Antheraea yamamai Bombyx mori evolution fibroin silk transgenic

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Rescue of white egg 1 mutant by introduction of the wildtype Bombyx kynurenine 3-monooxygenase gene

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Insect Science 2007年第2期14卷 85-92页

作者： GUO-XING QUAN ISAO KOBAYASHI KATSURA KOJIMA keiro uchino TOSHIO KANDA HIDEKI SEZUTSU TORU SHIMADA TOSHIKI TAMURA Transgenic Silkworm Research Center National Institute of Agrobiological Sciences Tsukuba Ibaraki 305-8634 Japan Biomimetic Laboratory National Institute of Agrobiological Sciences Tsukuba Ibaraki 305-8634 Japan Department of Agricultural and Environmental Biology Graduate School of Agricultural and Life Sciences The University of Tokyo Tokyo Japan

In silkworms, the white egg 1 （w-1） mutant, which is characterized by white eyes and white eggs, is deficient in Bombyx kynurenine 3-monooxygenase （KMO） activity. To investigate whether the w-1 mutant phenotype is rescued by introducing the wild-type KMO gene, we constructed transgenic silkworms with the wild-type Bombyx KMO gene under the control of either the cytoplasmic actin gene promoter （A3KMO） or the native KMO gene promoter （KKMO）. We created two transgenic lines with A3KMO and one line with KKMO constructs. The eyes of adults in these lines were brown, and the eggs laid by the transgenic females were also brown. Reverse transcription-polymerase chain reaction（RT-PCR） analysis showed that the A3KMO silkworm lines expressed the transcript in the mid-gut, fat bodies, and Malpighian tubules. The KKMO line expressed the transcript only in the fat bodies and Malpighian tubules. The intensity of eye and egg color in the transgenic lines was proportional to the KMO expression level. Interestingly, transgenic larvae with the A3KMO construct had a light brown larval cuticle, but the KKMO line did not. These results indicate that the wild-type KMO gene can be used as a marker gene for visually screening transgenic silkworms.

关键词： Bombyx mori KMO marker gene transgenic w-1

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