멧누에(Bombyx mandarina)로부터 Chitinase를 코딩하는 cDNA의 분리 및 염기서열 결정

Molecular Cloning and Characterization of the Gene Encoding Chitinase from Bombyx mandarina

  • 구태원 (농촌진흥청 농업과학기술원) ;
  • 황재삼 (농촌진흥청 농업과학기술원) ;
  • 성규병 (농촌진흥청 농업과학기술원) ;
  • 윤은영 (농촌진흥청 농업과학기술원) ;
  • 방혜선 (농촌진흥청 농업과학기술원) ;
  • 권오유 (충남대학교 의과대학)
  • 발행 : 1999.08.01

초록

Insects use chitinolytic enzyme to digest chitin in the exoskelton during the molting process. We have isolated and sequenced a chitinase-encoding cDNA from the silkworm, Bombyx mandarina, compared its sequenced with genes encoding chitinolytic enzymes from other sources. The insert DNA in the clone is 2,675 nucleotides long with an open reading frame of 1,695 uncletides that encodes a protein of 565 amino acids with a molecuar weight of 63.4 kDa. The 3' -untranslated region of 889 nucleotides is AT-rich and contains two putative polyadenylation signals. The N-terminal sequence of the encoded protein contains numerous hydrophobic residues characteristic of a leader peptide. The amino acid alignment revealed that the endo-$\beta$-N-acetylglucosaminidase had 83% and 97% homology with M. sexta and B. mori, respectively. The deduced amino acid had two highly conserved region at the amino acid residues 97-111 and 139-148 that were related to the existing chitinase.

키워드

참고문헌

  1. Insect Biochem v.17 Effect of 20-hydoxyecdysone on chitinase and β-N-acetylglucosaminidase during the larval-pupal transformation of Manduca sexta(L.) Fukamizo, T.;Kramer, K. J.
  2. Insect Biochem v.25 Baculovirus-mediated expression of a Manduca sexta chitinase gene;properties of the recombinant protein Gopalakrishan, B.;Muthukrishnan, S.;Kramer, K. J
  3. Prot. seq. data. Anal. v.3 Weak sequence homologies among chitinases detected by clustering analysis Henrissat, B.
  4. EMBO J. v.5 Isolation and charaterization of the genes encoding two chitinase enzymes from Serratia marcescens Jones, F. S. G.;Grady, K. L.;Suslow, T. VC.;Bedlbreek, J. R.
  5. Insect Biochem. Mol. Biol. v.28 no.3 Molecular cloning of chitinase cDNAs from the silkworm, Bomyx mori and the fall webworm, Hyphantria cunea Kim, M. G.;Shin, S. W.;Bae, K. S.;Park, H. Y.
  6. Insect Biochem v.19 Apperance of chitinolytic enzymes in integument of Bombyx mori during the larval-pupal transformation Koga, D.;Fujimoto, H.;Funakoshi, T.;Utsumi, T.;Ide,ㅁ.
  7. Insect Biochem. v.13 Insect endochitinase : glycoproteins from molting fluied, integument and pupal hemolymph of Manduca sexta L. Koga, D.;Jika, J.;Kramer, K. J.
  8. Insect Biochem. v.16 Insect chitin. Physical state, synthesis, degradation and metabolic regulation Kramer, K. J.;Koga, D.
  9. Biochemistry and Pharmacology v.3 Chitin metabolism in insects In Comprehensive Insect Physiology Kramer, K. J.;Dzisdik-Turner, C.;Koga, D.;Kerkut G. A.(ed.);Gilbert L. I.(ed.)
  10. Biochem. Mol. Biol. v.23 no.6 Sequnce of a cDNA and Expression of gene encoding epidermal and gut chitinase of Manduca sexta Insect Kramer, K. J.;Corpuz, L.;Choi, H. K.
  11. J. Biol. Chem. v.266 Chitinase is required for cell seperation during growth of Saccharomyces cerevisiae Kuranda, M. J.;Robbins, P. W.
  12. Proc. Natl. Acad. Sci. U. S. A. v.86 Isolation of complementry DNA encoding a chitinase with structural homology to a bifunctional lysozyme/chitinase Metraux, J. P.;Burkhart, W.;Moyer, M.;Dincheer, S.;Middlesteadt, W.;Willam, S.;Payne, G.;Carnes, M.;Ryals, J.
  13. Protein Eng. v.10 Indentification of prokaryotic and eukaryotic signal peptides and prediction of their cleavage site Nielsen, H.
  14. Archs Insect Biochem. Physiol v.2 Synthesis and deposition of chitin in larvae of Australian sheep blowfly, Lucilia cuprina Ratnakaran, A.;Hackman, R. H.
  15. J. Bio. CHem. v.259 Primary structure of the Streptomyces enzyme Endo-β-N-acetylglucosaminidase H. Robbins, P. W.;Trimble, R. B.;Wirth, D. F.;Herring, C.;Maley, F.;Maley, G. F.;Das, R.;Gibson, B. W.;Royal, N.;Biemann, K.
  16. Molecular cloning. A Laboratory Manual(2nd ed.) Sambook, J.
  17. Structure v.2 Crystal structures of hevamine, a plant defence protein with chitinase and lysozyme activity, and its complex with an inhibitor Van Scheltinga, A. C. T.;Kalk, K. H. Beintema, I. J.;Dijkstra, B. W.
  18. Molecular Ecology and Evolution : Approaches and Applications Evolution and multifuntionality of the chitin system Wagner, G. P.;Schierwater B.(ed.);Streit B.(ed.);Wagner G. P.(ed.);DeSalle, R.(ed.)
  19. J. Bacteriol v.174 Structure of the gene encoding chitinase D of Bacillus circuland WL-12 and possible homology of the enzyme to other prokaryotic chitinases and classIII plant chitinases Wanatbe, T.;Oyanagi, W.;Suzuki, K.;Ohnishi, K.;Tanaka, H.