• Title/Summary/Keyword: Transgenic silkworm

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Cloning of the posterior silk glands specific-expressed gene of silkworm (누에 후부실샘 특이 발현 유전자 클로닝)

  • Piao, Yulan;Kim, Seong-Ryul;Kim, Sung-Wan;Kang, Seok-Woo;Goo, Tae-Won;Choi, Kwang-Ho
    • Journal of Sericultural and Entomological Science
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    • v.53 no.1
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    • pp.44-49
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    • 2015
  • We characterized tissue specific-expressed genes in the posterior silk gland of Bombyx mori using by the Annealing Control Primer based differential display-PCR manner. In this study, we isolated 34 differentially expressed PCR amplicons, which one of these was identified as a novel transcript named as ACP-16 (366 bp), its expression was observed only in the posterior silk glands by Northern blot analysis. To determine promoter region of the ACP-16, we isolated and analyzed a phage DNA having 1.7 kb-long genome DNA including the open reading flame and 5'- upstream untranslated region of the ACP-16 gene from a genomic DNA library. We have estimated a promoter region of the ACP-16 gene by a web promoter prediction engine, which locates -750 ~ -165 from translation initiation site (ATG, +1). ACP-16 gene is necessary to more studies about critical biological role in order to apply the silkworm's transgenic system.

Characterization of the Promoter Controling the Stage-Specific Gene Expression of Bombyx mori (누에를 이용한 시기 특이적 발현 조절 유전자 promoter 개발)

  • Park, Seung-Won;Choi, Gwang-Ho;Goo, Tae-Won;Kim, Seong-Ryul;Kang, Seok-Woo
    • Journal of Life Science
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    • v.21 no.10
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    • pp.1466-1472
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    • 2011
  • We characterized embryo early gene (EEG)-704 promoter of the silkworm Bombyx mori, which is specifically regulated in the development stages. To determine core promoter region, 10 different partial mutant clones were tested by luciferase assay in Sf9 cells. About 1.5 kb promoter shows higher luciferase activity than constitutive promoter (BmA3). Interestingly, EEG-704 shares the same DNA sequences with BmHsp20.8 by the result of BLAST analysis; its expression is also increased under heat shock condition. Development of such promoter inducible, directly or indirectly in the developmental-stage, is very useful in making recombinant proteins in transgenic silkworms.

Bacillus thuringiensis as a Specific, Safe, and Effective Tool for Insect Pest Control

  • Roh, Jong-Yul;Choi, Jae-Young;Li, Ming-Sung;Jin, Byung-Rae;Je, Yeon-Ho
    • Journal of Microbiology and Biotechnology
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    • v.17 no.4
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    • pp.547-559
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    • 2007
  • Bacillus thuringiensis (Bt) was first described by Berliner [10] when he isolated a Bacillus species from the Mediterranean flour moth, Anagasta kuehniella, and named it after the province Thuringia in Germany where the infected moth was found. Although this was the first description under the name B. thuringiensis, it was not the first isolation. In 1901, a Japanese biologist, Ishiwata Shigetane, discovered a previously undescribed bacterium as the causative agent of a disease afflicting silkworms. Bt was originally considered a risk for silkworm rearing but it has become the heart of microbial insect control. The earliest commercial production began in France in 1938, under the name Sporeine [72]. A resurgence of interest in Bt has been attributed to Edward Steinhaus [105], who obtained a culture in 1942 and attracted attention to the potential of Bt through his subsequent studies. In 1956, T. Angus [3] demonstrated that the crystalline protein inclusions formed in the course of sporulation were responsible for the insecticidal action of Bt. By the early 1980's, Gonzalez et al. [48] revealed that the genes coding for crystal proteins were localized on transmissible plasmids, using a plasmid curing technique, and Schnepf and Whiteley [103] first cloned and characterized the genes coding for crystal proteins that had toxicity to larvae of the tobacco hornworm, from plasmid DNA of Bt subsp. kurstaki HD-1. This first cloning was followed quickly by the cloning of many other cry genes and eventually led to the development of Bt transgenic plants. In the 1980s, several scientists successively demonstrated that plants can be genetically engineered, and finally, Bt cotton reached the market in 1996 [104].

Reeling of recombinant flourescence cocoons through low temperature decompressed cooking (저온감압 자견법에 의한 재조합 형광누에고치의 조사)

  • Park, Jong-Hwa;Kim, Sung-Wan;Jeong, Young-Hun;Lee, Jong-Kil;Go, Young-Mi;Lee, Sang-Chan;Choi, Kwang-Ho;Kim, Seong-Ryul;Goo, Tae-Won
    • Journal of Sericultural and Entomological Science
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    • v.51 no.2
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    • pp.142-146
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    • 2013
  • The fluorescent proteins are generally denatured by heat treatment and thus lose their color. The normal reeling method includes processing by drying and cooking the cocoons near $100^{\circ}C$ before reeling. Therefore, the usual processing method cannot be used for making colored fluorescent silks. To develop a method that is applicable to producing transgenic silk without color loss, we develop reeling methods adequate for a recombinant fluorescence cocoons. It was found that the fluorescence cocoons keep their native color when dried at temperatures lower than $60^{\circ}C$ for 15 h. Also, a new cooking method to soften the fluorescent cocoons was developed: the cocoons were soaked in a solution of 0.2% sodium carbonate ($Na_2CO_3$)/0.1% nonionic surfactant (Triton X100) at $60^{\circ}C$ and then placed under vacuum. The repeated vacuum treatments enabled complete penetration of the solution into the cocoons, and the cocoons were thus homogenously softened and ready for reeling. In this state, the cooked cocoons can be reeled by an automated reeling machine. Our results suggest that drying and cooking of the cocoons at low temperature enables the subsequent reeling of the colored fluorescent silks by an automatic reeling machine without color loss and can produce silks that can be used for making higher value-added silk materials.