International Journal of Industrial Entomology and Biomaterials

Korean Society of Sericultural Science (한국잠사학회)
권호 표지

Molecular Cloning and Characterization of the Fibroin Light Chain Gene from the Silkworm Baekok-Jam (Bombyx mori)

  • Park, Kwang-Ho (College of Natural Resources and Life Science, Dong-A University, Department of Sericulture and Entomology, National Institute of Agricultural Science & Technology, RDA) ;
  • Goo, Tae-Won (Department of Sericulture and Entomology, National Institute of Agricultural Science & Technology, RDA) ;
  • Yun, Eun-Young (Department of Sericulture and Entomology, National Institute of Agricultural Science & Technology, RDA) ;
  • Hwang, Jae-Sam (Department of Sericulture and Entomology, National Institute of Agricultural Science & Technology, RDA) ;
  • Kang, Seok-Woo (Department of Sericulture and Entomology, National Institute of Agricultural Science & Technology, RDA) ;
  • Lee, Sang-Mong (Department of Sericultural and Entomological Biology, Miryang National University) ;
  • Sohn, Hung-Dae (College of Natural Resources and Life Science, Dong-A University) ;
  • Jin, Byung-Rae (College of Natural Resources and Life Science, Dong-A University)
  • Published : 2002.09.01
Download PDF
⟨ Previous Next ⟩

Abstract

We have cloned and characterized the complete fibroin light chain gene from the silkworm Baekok-Jam, Bombyx mori, a recommended variety in Korea. It consists of seven exons and six introns. It consists of 14,663 nucleotides long with an open reading frame of 786 nucleotides that encodes a protein of 262 amino acid residues with a molecular mass of approximately 26,000 dalton. The amino acid alignment revealed that the Baekok-Jam fibroin light chain had 98.5 % protein sequence identity to J139 strain: differed at four amino acid positions (11, 46, 80 and 123). The Northern hybridization analysis showed that the Baekok-Jam fibroin light chain gene was specifically expressed in the posterior silk gland.

Keywords

References

  1. Breathnach, R., C. Benoist, K. OHara, F. Gannon and P. Cham-bon (1978) Ovalbumin gene: evidence for a leader sequence in the mRNA and DNA sequences at the exon-intron bound-aries. Proc. Natl. Acad. Sci. USA 75, 4853-4857 https://doi.org/10.1073/pnas.75.10.4853
  2. Corden, J., B. Wasylyk, A. Buchwalder, P. Sassone-Corsi, C. Kedinger and P. Chambon (1980) Promoter sequences of eukaryotic protein-coding genes. Science 209, 1406-1414 https://doi.org/10.1126/science.6251548
  3. Hui, C. C., K. Matsuno and Y. Suzuki (1990a) Fibroin gene promoter contains a cluster of homeodomain binding sites that interact with three silk gland factors. J. Mol. Biol. 20, 651-670
  4. Hui, C. C., Y. Suzuki, Y. Kikuchi and S. Mizuno (1990b) Homeodomain binding sites in the 5' flanking region of the Bombyx mori silk fibroin light-chain gene. J. Mol. Biol. 5 395-398
  5. Inoue, S., K. Tanaka, F. Arisaks, S. Kimura, K. Ohtomo and S. Mizuno (2000) Silk fibroin of Bombyx mori is secreted, assembling a high molecular mass elementary unit consist-ing of H-chain, L-chain and P25, with a 6:6:1 molar ratio. J. BioI. Chem. 275, 40517-40528 https://doi.org/10.1074/jbc.M006897200
  6. Kikuchi, Y., K. Moh, S. Suzuki, K. Yamaguchi and S. Mizuno (1992) Structure of the Bombyx mori fibroin light-chain-encoding gene: upstream sequence elements common to the light and heavy chain. Gene 15, 151-158 https://doi.org/10.1016/0378-1119(81)90124-4
  7. Kimura, K., F. Oyama, H. Ueda, S. Mizuno and K. Shimura (1985) Molecular cloning of the fibroin light chain comple-mentary DNA and its use in the study of the expression of the light chain gene in the posterior silk gland of Bombyx mori. Experientia 41, 1167-1171 https://doi.org/10.1007/BF01951711
  8. McMaster, G. K. and G. G. Carmichael (1977) Analysis of sin-gle- and double-stranded nucleic acids on polyacryamide and agarose gels by using glyoxal and acridine orange. Proc. Natl. Acad. Sci. USA 74, 4835-4838 https://doi.org/10.1073/pnas.74.11.4835
  9. Mori, H., M. Yamao, H. Nakazawa, Y. Sugahara, N. Shirai, F. Matsubara, M. Sumida and T. Imamura (1995) Transovarian transmission of a foreign gene in the silkworm, Bombyx mori, By Autographa califomica nuclear polyhedrosis virus. BioTchnoloey 13, 1005-1007 https://doi.org/10.1038/nbt0995-1005
  10. Proudfoot, N. J. and G. G. Brownlee (1976) Non-coding region sequences in eukaryotic messenger RNA. Nature 263, 211-214 https://doi.org/10.1038/263211a0
  11. Shapiro, M. B. and P. Senapathy (1987) RNA splice junctions of different classes of eukaryotes: sequence statistics and functional implications in gene expression. Nucleic Acids Res.15,7155-7174 https://doi.org/10.1093/nar/15.17.7155
  12. Takei, F., Y. Kikuchi, A. Kikuchi, S. Mizuno and K. Shimura (1987) Further evidence for impotence of the subunit combi-nation of silk fibroin polypeptides in its efficient secretion from the posterior silk gland cells. J. Cell Biol. 105,175-180 https://doi.org/10.1083/jcb.105.1.175
  13. Tanaka, K., K. Mori and S. Mizuno (1993) Immunological identification of the major disulfide-linked light component of silk fibroin. J. Biochem. 114, 1-4 https://doi.org/10.1093/oxfordjournals.jbchem.a124122
  14. Tanaka, K., S. Inoue and S. Mizuno (1999) Hydrophobic inter-action of P25, containing Asn-linked oligosaccharide chains, with the H-L complex of silk fibroin produced by Bombyx mori. Insect Biochem. Mol. Biol. 29, 269-276 https://doi.org/10.1016/S0965-1748(98)00135-0
  15. Yamaguchi, K., Y. Kikuchi, T. Takagi, A. Kikuchi, F. Oyama, K. Shimura and S. Mizuno (1989) Primary structure of the silk fibroin light chain determined by cDNA sequencing and peptide analysis. J. Mol. Biol. 210, 127-139 https://doi.org/10.1016/0022-2836(89)90295-7
  16. Zhou, C. Z., P. Confalonieri, N. Medina, Y. Zivanovic, C. Esnault, T. Yang, M. Jacquet, J. Janin, M. Duguet, R. Per-asso. and Z. G. Li (2000) Fine organization of Bombyx mori fibroin heavy chain gene. Nucleic Acids Res. 28, 2413-2419 https://doi.org/10.1093/nar/28.12.2413