Bombyx mori b-tubulin Promoter for High-level Expression of Heterologous Genes
2
조회
0
다운로드
0
인용
0
참고문헌
키워드
초록
We previously isolated 9 clones that show stronger signal compared to Bombyx mori cytoplasmic actin gene (BmA3) by using a dot blot hybridization. In this study, we focused on one clone among these clones which has high amino acid similarity with b-tubulin gene of B. mori. This clone was ubiquitously expressed in all tissues and developmental stage of B. mori. As result of promoter assay using dual luciferase assay system, we found the highest transcription activity region (-750/-1) in the 5’-flanking region of b-tubulin gene, which has about 47 fold more intensive promoter activity than BmA3 promoter. Moreover, the b-tubulin promoter was normally regulated in Bm5, Sf9, and S2 cells. Therefore, we suggest that b-tubulin promoter may be used more powerful and effectively for transgene expression in various insects containing B. mori as a universal promoter.
Abstract
We previously isolated 9 clones that show stronger signal compared to Bombyx mori cytoplasmic actin gene (BmA3) by using a dot blot hybridization. In this study, we focused on one clone among these clones which has high amino acid similarity with b-tubulin gene of B. mori. This clone was ubiquitously expressed in all tissues and developmental stage of B. mori. As result of promoter assay using dual luciferase assay system, we found the highest transcription activity region (-750/-1) in the 5’-flanking region of b-tubulin gene, which has about 47 fold more intensive promoter activity than BmA3 promoter. Moreover, the b-tubulin promoter was normally regulated in Bm5, Sf9, and S2 cells. Therefore, we suggest that b-tubulin promoter may be used more powerful and effectively for transgene expression in various insects containing B. mori as a universal promoter.
참고문헌 (24)
- Ailor E, "Modifying secretion and posttranslational processing in insect cells", Curr Opin Biotechnol, 10, pp.142~145 (1999)
- 구태원, "Utilization of the Bombyx mori Hypothetical Protein 32 Promoter for Efficient Transgene Expression", International Journal of Industrial Entomology and Biomaterials, 20(2), pp.107~114 (2010), 한국잠사학회
- 구태원, "A Powerful Ubiquitous Activity of Bombyx mori Heat Shock Protein 70 Promoter", Genes & Genomics, 33(6), pp.635~643 (2011), 한국유전학회
- Handler AM, "The lepidopteran transposon vector, piggyBac, mediates germline transformation in the Mediterranean fruit fly", Proc Natl Acad Sci USA, 9513, pp.7520~7525 (1998)
- Horn C, "Fluorescent transformation markers for insect transgenesis", Insect Biochem Mol Biol, 32, pp.1221~1235 (2002)
- Jacobs PP, "N-glycosylation engineering of biopharmaceutical expression systems", Curr Mol Med, 9, pp.774~800 (2009)
- Kato T, "Silkworm expression system as a platform technology in life science", Appl Microbiol Biotechnol, 85, pp.459~470 (2010)
- Löwe J, "Crystal structure of the bacterial cell-division protein FtsZ", Nature, 8, pp.203~206 (1998)
- Mangé A, "A strong inhibitory element down-regulates SRE-stimulated transcription of the A3 cytoplasmic actin gene of Bombyx mori", J Mol Biol, 265, pp.266~274 (1997)
- Nogales E, "Tubulin and FtsZ form a distinct family of GTPases", Nat Struct Biol, 5, pp.451~458 (1998)
- Qian Q, "Highefficiency production of human serum albumin in the posterior silk glands of transgenic silkworms, Bombyx mori L", PLoS One, 13, pp.e0191507~ (2018)
- Royer C, "Biosynthesis and cocoon export of a recombinant globular protein in transgenic silkworms", Transgenic Res, 14, pp.463~472 (2005)
- A methods for baculovirus vector and insect cell culture procedures, "Summers MD", Texas Agricultural Experiment Station, ~, pp.Bulletin No. 1555 (1987)
- Sun SC, "Hemolin : an insect-immune protein belonging to the immunoglobulin superfamily", Science, 250, pp.1729~1732 (1990)
- Tamura T, "Germline transformation of the silkworm Bombyx mori L. using a piggyBac transposon-derived vector", Nat Biotechnol, 18, pp.81~84 (2000)
- Thomas JL, "3xP3-EGFP marker facilitates screening for transgenic silkworm Bombyx mori L. from the embryonic stage onwards", Insect Biochem Mol Biol, 32, pp.247~253 (2002)
- Tomita M, "Transgenic silkworms produce recombinant human type III procollagen in cocoons", Nat Biotechnol, 21, pp.52~56 (2003)
- Tomita M, "A germline transgenic silkworm that secretes recombinant proteins in the sericin layer of cocoon", Transgenic Res, 16, pp.449~465 (2007)
- Tryselius Y, "CecC, a cecropin gene expressed during metamorphosis in Drosophila pupae", Eur J Biochem, 204, pp.395~399 (1992)
- Uchino K, "Germ line transformation of the silkworm, Bombyx mori, using the transposable element Minos. Mol. Genet", Genomics, 277, pp.213~220 (2007)
- Uhlírová M, "Heat inducible transgenic expression in the silkmoth Bombyx mori", Dev Genes Evol, 212, pp.145~151 (2002)
- Feng Wang, "Large-scale production of bioactive recombinant human acidic fibroblast growth factor in transgenic silkworm cocoons", Scientific Reports, 5(1) (2015), Springer Science and Business Media LLC[DOI]
- Wurm FM, "Human therapeutic proteins from silkworms", Nat. Biotechnol, 21, pp.34~35 (2003)
- Zhao A, "New and highly efficient expression systems for expressing selectively foreign protein in the silk glands of transgenic silkworm", Transgenic Res, 19, pp.29~44 (2010)