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Korean Society for Biochemistry and Molecular Biology (생화학분자생물학회)
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Isolation and Characterization of Major Royal Jelly cDNAs and Proteins of the Honey Bee (Apis cerana)

  • Srisuparbh, Duangporn (Department of Biochemistry, Faculty of Science, Chulalongkorn University) ;
  • Klinbunga, Sirawut (National Center for Genetic Engineering and Biotechnology (BIOTEC), National Science and Technology Development Agency (NSTDA)) ;
  • Wongsiri, Siriwat (Department of Biology, Faculty of Science, Chulalongkorn University) ;
  • Sittipraneed, Siriporn (Department of Biochemistry, Faculty of Science, Chulalongkorn University)
  • Received : 2003.03.17
  • Accepted : 2003.06.04
  • Published : 2003.11.30
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Abstract

An expressed sequence tag (EST) library was established from the hypopharyngeal glands of Apis cerana. Sixty-six recombinant clones, possessing inserts >500 bp, were randomly selected and unidirectional sequenced. Forty-two of these (63.6%) were identified as homologues of Major Royal Jelly Proteins families 1, 2, 3, and 4 of A. mellifera (AmMRJP) for which MRJP1 was the most abundant family. The open-reading frame of the MRJP1 homologue (AcMRJP1) was 1299 nucleotides that encoded 433 deduced amino acids with three predicted N-linked glycosylation sites. The AcMRJP1 sequence showed 93% and 90% homologies with nucleotide and deduced amino acid sequences of AmMRJP1, respectively. Two complete transcripts of apisimin, and one and two partial transcripts of $\alpha$-glucosidase and glucose oxidase, were also isolated. In addition, the royal jelly proteins of A. cerana were purified and characterized using Q-Sepharose and Sephadex G-200 column chromatography. The native forms of protein peaks A1, A2, B1, and C1 were 115, 55, 50, and 300 kDa, respectively. SDS-PAGE analysis indicated that A1 and C1 were dimeric and oligomeric forms of the 80 kDa and 50 kDa subunits, respectively. The ratio of the total protein quantities of A1 : A2 : B1 : C1 were 2.52 : 4.72 : 1 : 12.21. Further characterization of each protein, using N-terminal and internal peptide sequencing, revealed that the respective proteins were homologues of MRJP3, MRJP2, MRJP1, and MRJP1 of A. mellifera.

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References

  1. Bilikova, K., Hanes, J., Nordhoff, E., Saenger, W., Klaudiny, J.and Simuth, J. (2002) Apisimin, a new serine-valine-richpeptide from honeybee (Apis mellifera L.) royal jelly:purification and molecular characterization. FEBS Lett. 528,125-129. https://doi.org/10.1016/S0014-5793(02)03272-6
  2. Bilikova, K., Klaudiny, J. and Simuth, J. (1999) Characterizationof the basic major royal jelly protein MRJP2 of honeybee(Apis mellifera) and its preparation by heterologous expressionin E.coli. Biologia, Bratislava 54, 733-739.
  3. Brouwers, E. V. M. (1982) Measurement of hypopharyngeal glandactivity in the honeybee. J. Apic. Res. 21, 193-198.
  4. Fukuda, M. and Kobata, A. (1993) Glycobiology, Oxforduniversity Press, New York, USA.
  5. Hanes, J. and Simuth, J. (1992) Identification and partialcharacterization of the major royal jelly protein of the honeybee (Apis mellifera L.). J. Apic. Res. 31, 22-26.
  6. Howe, S. E., Dimick, P. S. and Benton, A. W. (1985)Composition of freshly harvested and commercial royal jelly. J.Apic. Res. 24, 52-61.
  7. Huang, Z. Y., Otis, G. W. and Teal, P. E. A. (1989) Nature ofbrood signal activating the protein synthesis of hypopharyngealgland in honey bees, Apis mellifera (Apidae: Hymenoptera).Apidologie 20, 455-464. https://doi.org/10.1051/apido:19890601
  8. Judova, J., Klaudiny, J. and Simuth, J. (1998) Preparation ofrecombinant most abundant MRJP1 of royal jelly. Biologia,Bratislava 53, 777-784.
  9. Karaali, A., Meydanoglu, F. and Eke, D. (1988) Studies oncomposition, freeze-drying and storage of Turkish royal jelly. J.Apic. Res. 27, 182-185.
  10. Kimura, M. (1980) A simple method for estimating evolutionaryrate of base substitutions through comparative studies ofnucleotide sequences. J. Mol. Evol. 16, 111-120. https://doi.org/10.1007/BF01731581
  11. Klaudiny, J., Hanes, J., Kulifajova, J., Albert, S. and Simuth, J.(1994) Molecular cloning of two cDNAs from the head of thenurse honey bee (Apis mellifera L.) for coding related proteinsof royal jelly. J. Apic. Res. 33, 105-111.
  12. Knecht, D. and Kaatz, H. H. (1990) Patterns of larval foodproduction by hypopharyngeal glands in adult worker honeybees. Apidologie 21, 457-468. https://doi.org/10.1051/apido:19900507
  13. Kubo, T., Sasaki, M., Nakamura, J., Sasagawa, H., Ohashi, K.,Takeuchi, H. and Natori, S. (1996) Change in the expression ofhypopharyngeal-gland/proteins of the worker honeybee (Apismellifera L.) with age and/or role. J. Biochem. 119, 291-295. https://doi.org/10.1093/oxfordjournals.jbchem.a021237
  14. Laemmli, U. K. (1970) Cleavage of structural proteins during theassembly of the head of bacteriophage T4. Nature 227, 680-685. https://doi.org/10.1038/227680a0
  15. Lensky, L. and Rakover, Y. (1983) Separate protein bodycompartments of worker honeybee (Apis mellifera L.) Comp.Biochem. Physiol. 75B, 607-615.
  16. Lowry, O. H., Rosebrough, N. J. Farr, A. L. and Randall, R. J.(1951) Protein measurement with the folin phenol reagent. J.Biol. Chem. 193, 265-275.
  17. Maniatis, T., Sambrook, J. and Fritsch, E. F. (1982) MolecularCloning: A Laboratory Manual. Cold Spring HarborLaboratory, Cold Spring Harbor Press, New York, USA.
  18. Ohashi, K., Natori, S. and Kubo, T. (1997) Change in the mode ofgene expression of the hypopharyngeal gland cells with an age-dependentrole change of the worker honeybee Apis melliferaL. Eur. J. Biochem. 249, 797-802. https://doi.org/10.1111/j.1432-1033.1997.t01-1-00797.x
  19. Ohashi, K., Natori, S. and Kubo, T. (1999) Expression of amylaseand glucose oxidase in the hypopharyngeal gland with an age-dependentrole change of the worker honeybee (Apis melliferaL.) Eur. J. Biochem. 265, 127-133. https://doi.org/10.1046/j.1432-1327.1999.00696.x
  20. Palma, M. S. (1992) Composition of freshly harvested Brazilianroyal jelly: identification of carbohydrates from the sugarfraction. J. Apic. Res. 31, 42-44.
  21. Pothichot, S. and Wongsiri, S. (1993). Attempts in queen rearingof Apis cerana larvae in Apis mellifera colonies and Apismellifera larvae in Apis cerana colonies. Asian Apicul. 128-133.
  22. Schmitzova, J., Klaudiny, J., Albert, S., Schroder, W.,Schreckengost, W., Hanes, J., Judova, J. and Simuth, J. (1998)A family of major royal jelly proteins of the honeybee Apismellifera L. Cell. Mol. Life Sci. 54, 1020-1030. https://doi.org/10.1007/s000180050229
  23. Simuth, J. (2001) Some properties of the main protein of honey bee (Apis mellifera) royal jelly. Apidologie 32, 69-80. https://doi.org/10.1051/apido:2001112
  24. Takenaka, T. and Takenaka, Y. (1996) Royal jelly from Apiscerana japonica and Apis mellifera. Biosci. Biotech. Biochem.60, 518-520. https://doi.org/10.1271/bbb.60.518
  25. Tomada, G., Matsuyama, J. and Matsuka, M. (1977) Studies onprotein in royal jelly. 2. Fractionation of water-soluble protein by DEAE-cellulose chromatography, gel filtration and discelectrophoresis. J. Apic. Res. 16, 125-130.
  26. Towbin, H., Starhelin, T. and Gordon, J. (1979) Electrophoretictransfer of proteins from polyacrylamide gels to nitrocellulosesheets: Procedure and some applications. Proc. Natl. Acad. Sci.USA 76, 4350-4354. https://doi.org/10.1073/pnas.76.9.4350

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