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Seong Wan Kim,Seon Young Kim,Hye Lim Yeo,Eun Young Yun,Kwang-Ho Choi,Seong Ryul Kim,Seok Woo Kang,Seung Won Park,Tae Won Goo 한국응용곤충학회 2013 한국응용곤충학회 학술대회논문집 Vol.2013 No.10
Immune-inducible antimicrobial peptides were produced using transgenic silkworms that expressed Rel family transcription factor, truncated BmRelish1 (BmRelish1t) genes under the control of the BmA3 promoter using the piggyBac vector. BmRelish1t gene contains all domains of Bmrelish: a Rel homolog domain (RHD), nuclear localization signal (NLS), acidic and hydrophobic amino acids (AHAA) rich region except the Ankyrin repeat domain (ANK) and the death domain (DD). (1:1) Mixtures of the donor vector (pG-3xP3EGFP-BmA3BmRelish1t) and helper vector were micro-injected into 1,800 eggs of bivoltin silkworms, Baegokjam and EGFP-induced fluorescence was observed for 25 broods of transgenic silkworms under a florescence stereomicroscope. Analysis by real-time PCR indicated that transgenic silkworms expressing BmRelish1t recombinant proteins displayed higher mRNA expression levels of the Bombyx mori antimicrobial peptides such as lebocin, moricin, and nuecin than the normal silkworms. Moreover, transgenic silkworms expressing BmRelish1t showed antibacterial activity against Escherichia coli. We suggest that transgenic expression of BmRelish1t may find useful applications for the production of various antimicrobial peptides at the same time in transgenic silkworms.
Utilization of the B. mori Heat Shock Protein 70 Promoter for Screening Transgenic Silkworms
Seong Wan Kim,Hye Lim Yeo,Seon Young Kim,Eun Young Yun,Kwang-Ho Choi,Seong Ryul Kim,Seok Woo Kang,Seung Won Park,Tae Won Goo 한국응용곤충학회 2013 한국응용곤충학회 학술대회논문집 Vol.2013 No.10
Silkworm transgenesis is now a routine method leading to a satisfactory yield of transformed animals and the reliable expression of transgenes during multiple successive generations. However, the screening of G1 transgenic individuals from numerous progeny has proved to be difficult and time-consuming work. Previously, we characterized the promoter of heat shock protein 70 from Bombyx mori (bHsp70), which is ubiquitously expressed in all tissues and developmental stages. To investigate the utilization of the bHsp70 promoter to screen transgenic individuals, the EGFP marker gene was inserted into the piggyBac vector under the control of the bHsp70 promoter. Mixtures of the donor and helper vectors were micro-injected into 3,060 eggs of bivoltine silkworms (Keomokjam). EGFP fluorescence was observed in 17 broods of transgenic silkworms under a florescence stereomicroscope. Interestingly, this fluorescent marker protein was detected not only in parts of the embryo segments on the seventh day of the G1 embryonic developmental stage but it was also detected in a part of the body of G1 hatched larvae, in the middle silk gland of G1 fifth instar larvae, and in the wings of seven-day-old G1 pupae and G1 moths. Therefore, we suggest that the bHsp70 promoter can be used for the rapid and simple screening of transgenic silkworms.
임성률(Lim, Seong Ryul),박순영(Park, Sun Yeong),곽천섭(Kwak, Chunsub),전인찬(Jeon, Inchan),최성종(Choi, Seong Jong) 한국방송·미디어공학회 2011 한국방송공학회 학술발표대회 논문집 Vol.2011 No.7
재난이 발생하였거나 발생할 우려가 있을 경우 국민들에게 이에 대한 정보를 신속하게 제공하면 많은 인명 및 재산 피해를 줄일 수 있을 것이다. 본 고에서는 우리나라에서 시행한 재난정보전달시스템에 대해 간략히 소개한다. 각 시스템의 장점 및 단점 또한 타 시스템과의 차별성에 대해 기술한다. 이러한 분석은 향후 국가통합재난정보전달시스템을 구축하기 위한 기본 자료로 활용될 수 있을 것이다.
Production of BmCecB1 antimicrobial peptide in the transgenic silkworm
Hye Lim Yeo,Seong Wan Kim,Seon Young Kim,Eun Young Yun,Kwang-Ho Choi,Seong Ryul Kim,Seok Woo Kang,Seung Won Park,Tae Won Goo 한국응용곤충학회 2013 한국응용곤충학회 학술대회논문집 Vol.2013 No.10
BmCecB1 are antimicrobial peptides from Bombyx mori and belongs to cecropin family. Antimicrobial peptides are important components of the innate immune systems in all living organism. This peptide has antibacterial activity against several Gram-positive and Gram-negative bacteria. To produce the BmCecB1 antimicrobial peptide, we constructed transgenic silkworm that expressed BmCecB1 gene under the control BmA3 promoter using piggyBac vector. The use of the 3xP3-driven EGFP cDNA as a marker allowed us to rapidly distinguish transgenic silkworm. Mixtures of the donor vector and helper vector were micro-injected into 600 eggs of bivoltin silkworms, Baegokjam. In total, 49 larvae (G0) were hatched and allowed to develop into moths. The resulting G1 generation consisted of 22 broods, and we selected 2 broods containing at least 1 EGFP-positive embryo. The rate of successful transgenesis for the G1 broods was 11%. We identified 9 EGFP-positive G1 moths and these were backcrossed with wild-type moths. With the aim of identifying a BmCecB1 as antimicrobial peptide, we investigated the Radical diffusion Assay (RDA) and then demonstrated that BmCecB1 possesses high antibacterial activities against Gram-negative bacteria.
Expression of the blue fluorescent protein (AmCyan) in the cocoon of transgenic silkworms
Seon Young Kim,Seong Wan Kim,Hye Lim Yeo,Eun Young Yun,Kwang-Ho Choi,Seong Ryul Kim,Seok Woo Kang,Seung Won Park,Tae Won Goo 한국응용곤충학회 2013 한국응용곤충학회 학술대회논문집 Vol.2013 No.10
To product the blue fluorescent protein (AmCyan) expressed cocoon, we were fused AmCyan cDNA to the heavy chain gene and injected the gene into a silkworm. AmCyan was one of the existing violet fluorochromes and originally derived from the fluorescent protein amFP486. AmFP486 was cloned from the sea anemone Anemonia majano (GenBank accession number AF168421), and belongs to the family of fluorescent proteins (FPs) isolated from coral reef organisms. The AmCyan fusion protein, each with N- and C- terminal sequences or the fibroin H-chain, were designed to be secreted into the lumen of the posterior silk glands. The expression of the AmCyan/H-chain fusion gene was regulated by the fibroin H-chain promoter. The use of the 3xP3 EGFP as a marker allowed us to rapidly distinguish transgenic silkworm. Mixtures of the donor and helper vectors were micro-injected into 300 eggs of bivoltine silkworms (Baegokjam). EGFP fluorescence was observed in 3 broods of transgenic silkworms under a florescence stereomicroscope. The cocoon was displayed strong blue fluorescence, proving that the fusion protein was present in the cocoon. Accordingly, we suggest that the AmCyan gene expressed cocoon will be enable the production of the novel biomaterials based on the transgenic silk.