Browse > Article

Utilization of the Bombyx mori Hypothetical Protein 32 Promoter for Efficient Transgene Expression  

Goo, Tae-Won (Department of Agricultural Biology, National Academy of Agricultural Science, RDA)
Kim, Sung-Wan (Department of Agricultural Biology, National Academy of Agricultural Science, RDA)
Kim, Seong-Ryul (Department of Agricultural Biology, National Academy of Agricultural Science, RDA)
Park, Seung-Won (Department of Agricultural Biology, National Academy of Agricultural Science, RDA)
Kang, Seok-Woo (Department of Agricultural Biology, National Academy of Agricultural Science, RDA)
Lee, Kwang-Gill (Department of Agricultural Biology, National Academy of Agricultural Science, RDA)
Kwon, O-Yu (Department of Anatomy, College of Medicine, Chungnam National University)
Yun, Eun-Young (Department of Agricultural Biology, National Academy of Agricultural Science, RDA)
Publication Information
International Journal of Industrial Entomology and Biomaterials / v.20, no.2, 2010 , pp. 107-114 More about this Journal
Abstract
For stable germline transformation, the promoter of Bombyx mori cytoplasmic actin gene (BmA3) has been used for ubiquitous expression of transgenes. So far, no strong promoter is available for ubiquitous expression in B. mori, excluding BmA3 promoter. To identify more powerful promoter than previously reported BmA3 promoter, we isolated 9 clones that show stronger signal compared to BmA3 by a dot blot hybridization. Among these 9 clones, we focused on one clone which has high amino acid homology (85%) with hypothetical protein 32 gene of Lonomia obliqua. This clone, named bHp32 (B. mori hypothetical protein 32) was ubiquitously expressed in all tissues and developmental stage of fifth instar B. mori larvae. As result of promoter assay using dual luciferase assay system, we found the highest transcription activity region (-1,200/+220) in the 5'-flanking region of bHp32 gene, which has 42-fold more intensive promoter activity than BmA3 promoter. Moreover, the bHp32 promoter was normally regulated in Bm5, Sf9, and S2 cells. Therefore, we suggest that bHp32 promoter may be used more powerful and effectively for transgene expression in various insects containing B. mori as a universal promoter.
Keywords
Bombyx mori; Hypothetical protein 32; Luciferase assay; Promoter;
Citations & Related Records
연도 인용수 순위
  • Reference
1 Uhlirova M, Asahina M, Riddiford LM, Jindra M (2002) Heatinducible transgenic expression in the silkmoth Bombyx mori. Dev Genes Evol 212, 145-151.   DOI   ScienceOn
2 Tamura T, Thibert C, Royer C, Kanda T, Abraham E, Kamba M, Komoto N, Thomas JL, Mauchamp B, Chavancy G, Shirk P, Fraser M, Prudhomme JC, Couble P (2000) Germline transformation of the silkworm Bombyx mori L. using a piggyBac transposon-derived vector. Nat Biotechnol 18, 81-84.   DOI   ScienceOn
3 Veiga AB, Ribeiro JM, Guimaraes JA, Francischetti IM (2005) A catalog for the transcripts from the venomous structures of the caterpillar Lonomia obliqua: identification of the proteins potentially involved in the coagulation disorder and hemorrhagic syndrome. Gene 355, 11-27.   DOI   ScienceOn
4 Wurm FM (2003) Human therapeutic proteins from silkworms. Nat Biotechnol 21, 34-35.   DOI   ScienceOn
5 Thomas JL, Da Rocha M, Besse A, Mauchamp B, Chavancy G (2002) 3xP3-EGFP marker facilitates screening for transgenic silkworm Bombyx mori L. from the embryonic stage onwards. Insect Biochem. Mol Biol 32, 247-253.
6 Zhao A, Zhao T, Zhang Y, Xia Q, Lu C, Zhou Z, Xiang Z, Nakagaki M (2010) New and highly efficient expression systems for expressing selectively foreign protein in the silk glands of transgenic silkworm. Transgenic Res 19, 29-44.   DOI   ScienceOn
7 Mange A, Julien E, Prudhomme JC, Couble P (1997) A strong inhibitory element down-regulates SRE-stimulated transcription of the A3 cytoplasmic actin gene of Bombyx mori. J Mol Biol 265, 266-274.   DOI   ScienceOn
8 Royer C, Jalabert A, Da Rocha M, Grenier AM, Mauchamp B, Couble P, Chavancy G (2005) Biosynthesis and cocoonexport of a recombinant globular protein in transgenic silkworms. Transgenic Res 14, 463-472.   DOI   ScienceOn
9 Summers MD, Smith GE (1987) A methods for baculovirus vector and insect cell culture procedures. Texas Agricultural Experiment Station, Bulletin No. 1555.
10 Sun SC, Lindstrom I, Boman HG, Faye I, Schmidt O (1990) Hemolin: an insect-immune protein belonging to the immunoglobulin superfamily. Science 250, 1729-1732.   DOI
11 Tomita M, Hino R, Ogawa S, Iizuka M, Adachi T, Shimizu K, Sotoshiro H, Yoshizato K (2007) A germline transgenic silkworm that secretes recombinant proteins in the sericin layer of cocoon. Transgenic Res 16, 449-465.   DOI   ScienceOn
12 Tomita M, Munetsuna H, Sato T, Adachi T, Hino R (2003) Transgenic silkworms produce recombinant human type III procollagen in cocoons. Nat Biotechnol 21, 52-56.   DOI   ScienceOn
13 Tryselius Y, Samakovlis C, Kimbrell DA, Hultmark D (1992) CecC, a cecropin gene expressed during metamorphosis in Drosophila pupae. Eur J Biochem 204, 395-399.   DOI   ScienceOn
14 Uchino K, Imamura M, Shimizu K, Kanda T, Tamura T (2007) Germ line transformation of the silkworm, Bombyx mori, using the transposable element Minos. Mol Genet Genomics 277, 213-220.   DOI   ScienceOn
15 Ailor E, Betenbaugh MJ (1999) Modifying secretion and posttranslational processing in insect cells. Curr Opin Biotechnol 10, 142-145.   DOI   ScienceOn
16 Horn C, Schmid BGM, Pogoda FS, Wimmer EA (2002) Fluorescent transformation markers for insect transgenesis. Insect Biochem Mol Biol 32, 1221-1235.   DOI   ScienceOn
17 Goo TW, Yun EY, Hwang JS, Kang SW, Park S, You KH, Kwon OY (2002) Molecular characterization of a Bombyx mori protein disulfide isomerase (bPDI). Cell Stress Chaperones 7, 118-125.   DOI   ScienceOn
18 Handler AM, McCombs SD, Fraser MJ Jr, Saul SH (1998) The Lepidopteran transposon vector, piggyBac, mediates germline transformation in the Mediterranean fruit fly. Proc Natl Acad Sci USA 9513, 7520-7525.
19 Hoog C (1991) Isolation of a large number of novel mammalian genes by a differential cDNA library screening strategy. Nucleic Acids Res 19, 6123-6127.   DOI   ScienceOn
20 Jacobs PP, Callewaert N (2009) N-glycosylation engineering of biopharmaceutical expression systems. Curr Mol Med 9, 774-800.   DOI   ScienceOn
21 Kato T, Kajikawa M, Maenaka K, Park EY (2010) Silkworm expression system as a platform technology in life science. Appl Microbiol Biotechnol 85, 459-470.   DOI   ScienceOn