BMB Reports

생화학분자생물학회 (Korean Society for Biochemistry and Molecular Biology)
권호 표지
DOI QR코드

DOI QR Code

Cloning, Expression and Genomic Organization of Genes Encoding Major Royal Jelly Protein 1 and 2 of the Honey Bee (Apis cerana)

  • Imjongjirak, Chanprapa (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)) ;
  • Sittipraneed, Siriporn (Department of Biochemistry, Faculty of Science, Chulalongkorn University)
  • 발행 : 2005.01.31
PDF 다운로드
⟨ 이전 논문 다음 논문 ⟩

초록

Major Royal Jelly Protein cDNAs of Apis cerana (AcMRJP) were cloned and characterized. The open reading frames (ORFs) of the AcMRJP1 and AcMRJP2 genes were 1302 and 1392 nucleotides, encoding 433 and 463 amino acid residues, respectively. The sequence divergences between AcMRJP1 and AcMRJP2 and their corresponding protein families in A. mellifera were 0.0618 and 0.0934 at the nucleotide level and 0.0912 and 0.1438 at the protein level, respectively. Phylogenetic analysis supports the orthologous similarity between these proteins. The deduced amino acids indicated high essential amino acid contents of AcMRJP1 and AcMRJP2 (47.5 and 44.8%, respectively). The genomic organization of both AcMRJP1 and AcMRJP2 was determined. Both the AcMRJP1 (3663 bp) and AcMRJP2 (3963 bp) genes contained six exons and five introns, where all boundaries conformed to the GT/AG rule. AcMRJP1 and AcMRJP2 cDNAs were cloned into pET17b, and both the recombinant (r) AcMRJP1 (47.9 kDa) and rAcMRJP2 (51.7 kDa) were expressed in the insoluble form. Western blot analysis and N-terminal sequencing of the solubilized proteins revealed successful expression of rAcMRJP1 and rAcMRJP2 in vitro. The yields of the purified rAcMRJP1 and rAcMRJP2 were approximately 20 and 8mg protein per liter of the flask culture, respectively.

키워드

참고문헌

  1. Albert, S., Bhattacharya, D., Klaudiny, J., Schmitzova and Simuth, J. (1999a) The family of major royal jelly proteins and its evolution. J. Mol. Evol. 49, 290-297 https://doi.org/10.1007/PL00006534
  2. Albert, S., Klaudiny, J. and Simuth, J. (1999b) Molecular characterization of MRJP3, highly polymorphic protein of honey bee (Apis mellifera) royal jelly. Insect Biochem. Mol. Biol. 29, 427-434 https://doi.org/10.1016/S0965-1748(99)00019-3
  3. Bilikova, K., Klaudiny, J. and Simuth, J. (1999) Characterization of the basic major royal jelly protein MRJP2 of honeybee (Apis mellifera) and its preparation by heterologous expression in E. coli. Biologia Bratislava 54, 733-739
  4. Felsenstein, J. (1993) Phylip (Phylogenetic Inference Package) Version 3.5c. Distributed by the author. Department of Genetics, University of Washington, Seattle, USA
  5. Hanes, J. and Simuth, J. (1992) Identification and partial characterization of the major royal jelly protein of the honey bee (Apis mellifera L.). J. Apic. Res. 31, 22-26
  6. Hirel, P. H., Schmitter, M. J., Dessen, P., Fayat, G. and Blanquet, S. (1989) Extent of N-terminal methionine excision from Escherichia coli proteins is governed by the sidechain length of the penultimate amino acid. Proc. Natl. Acad. Sci. USA 86, 8247-8251 https://doi.org/10.1073/pnas.86.21.8247
  7. Jones, G. and Sharp, P. A. (1997) Ultraspiracle: an invertebrate nuclear receptor for juvenile hormones. Proc. Natl. Acad. Sci. USA 94, 13499-13503 https://doi.org/10.1073/pnas.94.25.13499
  8. Judova, J., Klaudiny, J. and Simuth, J. (1998) Preparation of recombinant most abundant MRJP1 of royal jelly. Biologia Bratislava 53, 777-784
  9. Kimura, M. (1980) A simple method for estimating evolutionary rate of base substitutions through comparative studies of nucleotide sequences. J. Mol. Evol. 16, 111-120 https://doi.org/10.1007/BF01731581
  10. Klaudiny, J., Hanes, J., Kulifajova, J., Albert, S. and Simuth, J. (1994) Molecular cloning of two cDNAs from the head of the nurse honey bee (Apis mellifera L.) for coding related proteins of royal jelly. J. Apic. Res. 33, 105-111
  11. Knecht, D. and Kaatz, H. H. (1990) Patterns of larval food production by hypopharyngeal glands in adult worker honey bees. Apidologie 21, 457-468 https://doi.org/10.1051/apido:19900507
  12. Kubo, T., Sasaki, M., Nakamura, J., Sasagawa, H., Ohashi, K., Takeuchi, H. and Natori, S. (1996) Change in the expression of hypopharyngeal-gland/proteins of the worker honeybee (Apis mellifera L.) with age and/or role. J. Biochem. 119, 291-295 https://doi.org/10.1093/oxfordjournals.jbchem.a021237
  13. Laemmli, U. K. (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227, 680- 685 https://doi.org/10.1038/227680a0
  14. Lensky, L. and Rakover, Y. (1983) Separate protein body compartments of worker honeybee (Apis mellifera L.). Comp. Biochem. Physiol. 75, 607-615 https://doi.org/10.1016/0305-0491(83)90104-9
  15. Maniatis, T., Sambrook, J. and Fritsch, E. F. (1982) Construction of genomic libraries; in Molecular Cloning: A Laboratory Manual. pp. 269-309, Cold Spring Harbor Laboratory Press, New York, USA
  16. Malecova, B., Ramser, J., O'Brien, J. K, Janitz, M., Judova, J., Lehrach, H. and Simuth, J. (2003) Honeybee (Apis mellifera L.) mrjp gene family: computational analysis of putative promoters and genomic structure of mrjp1, the gene coding for the most abundant protein of larval food. Gene 303, 165-175 https://doi.org/10.1016/S0378-1119(02)01174-5
  17. Mitsuo, R., Sato, M. and Itakura, T. (2001) Cloning, sequencing and phylogenetic analysis of complementary DNA of novel cytochrome P-450 CYP1A in Japanese eel (Anguilla japonica). Mar. Biotechnol. 3, 218-223 https://doi.org/10.1007/s101260000070
  18. Okamoto, I., Taniguchi, Y., Kunikata, T., Kohno, K., Iwaki, K., Ikeda and M., Kurimoto, M. (2003) Major royal jelly protein 3 modulates immune responses in vitro and in vivo. Life Sci. 73, 2029-2045 https://doi.org/10.1016/S0024-3205(03)00562-9
  19. Pothichot, S. and Wongsiri, S. (1993) Attemps in queen rearing of Apis cerana larvae in Apis mellifera colonies and Apis mellifera larvae in Apis cerana colonies. Asian Apic. 128-133
  20. 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 Apis mellifera L. Cell. Mol. Life Sci. 54, 1020-1030 https://doi.org/10.1007/s000180050229
  21. Sergaves, W. A. (1991). Something old, some things new: the steroid receptor superfamily in Drosophila. Cell 67, 225-228 https://doi.org/10.1016/0092-8674(91)90172-U
  22. 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
  23. Smith, D. R. and Hagen, R. H. (1997) The biogeography of Apis cerana as revealed by mitochondrial DNA sequence data. J. Kansas Entomol. Soc. 64, 294-310
  24. Srisuparbh, D., Klinbunga, S., Wongsiri, S. and Sittipraneed, S. (2003) Isolation and characterization of major royal jelly cDNAs and proteins of the honey bee (Apis cerana). J. Biochem. Mol. Biol. 36, 572-579 https://doi.org/10.5483/BMBRep.2003.36.6.572
  25. Takenaka, T. and Takenaka, Y. (1996) Royal jelly from Apis cerana japonica and Apis mellifera. Biosci. Biotech. Biochem. 60, 518-520 https://doi.org/10.1271/bbb.60.518
  26. Thompson, J. D., Higgins, D. G. and Gibson, T. J. (1994) Clustal W: improving the sensitivity of progressive multiple sequence weighting, position-specific gap penalties and weight metric choices. Nucleic Acids Res. 22, 4673-4680 https://doi.org/10.1093/nar/22.22.4673
  27. Towbin, H., Starhelin, T. and Gordon, J. (1979) Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc. Natl. Acad. Sci. USA 76, 4350-4354 https://doi.org/10.1073/pnas.76.9.4350

피인용 문헌

  1. Antilipopolysaccharide factor (ALF) of mud crab Scylla paramamosain: Molecular cloning, genomic organization and the antimicrobial activity of its synthetic LPS binding domain vol.44, pp.12, 2007, https://doi.org/10.1016/j.molimm.2007.01.028
  2. Cloning, genomic organization and expression of two glycosyl hydrolase family 10 (GHF10) genes from golden apple snail (Pomacea canaliculata) vol.19, pp.3, 2008, https://doi.org/10.1080/10425170701517911
  3. Towards a molecular definition of worker sterility: differential gene expression and reproductive plasticity in honey bees vol.15, pp.5, 2006, https://doi.org/10.1111/j.1365-2583.2006.00678.x
  4. Recovery of major royal jelly protein 1 expressed inPichia pastorisin aqueous two-phase systems vol.89, pp.7, 2014, https://doi.org/10.1002/jctb.4342
  5. Identification, characterization and expression of adipose differentiation-related protein (ADRP) gene and protein in ovaries of the giant tiger shrimp Penaeus monodon vol.308, 2010, https://doi.org/10.1016/j.aquaculture.2010.06.039
  6. Molecular cloning and characterization of crustin from mud crab Scylla paramamosain vol.36, pp.5, 2009, https://doi.org/10.1007/s11033-008-9253-0
  7. Proteomic characterization of royal jelly proteins in Chinese (Apis cerana cerana) and European (Apis mellifera) honeybees vol.73, pp.6, 2008, https://doi.org/10.1134/S0006297908060072
  8. Economic analysis of royalactin production under uncertainty: Evaluating the effect of parameter optimization vol.31, pp.3, 2015, https://doi.org/10.1002/btpr.2073
  9. Isolation and characterization of progesterone receptor-related protein p23 (Pm-p23) differentially expressed during ovarian development of the giant tiger shrimp Penaeus monodon vol.308, 2010, https://doi.org/10.1016/j.aquaculture.2010.06.037
  10. Expression of Recombinant AccMRJP1 Protein from Royal Jelly of Chinese Honeybee in Pichia pastoris and Its Proliferation Activity in an Insect Cell Line vol.58, pp.16, 2010, https://doi.org/10.1021/jf1007133