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출처 : ncbi

Isolation and characterization of Major Royal Jelly cDNAS and proteins of

the Honey Bee(Apis cerana)

Duangporn Srisuparbh, Sirawut Klinbunga, Siriwat Wongsiri and Siriporn Sittipraneed

Department of Biochemistry, Department of Biology, Faculty of Science, Chulaongkom University, Bangkok 10330, Thailand

National Center for Genetic Engineering and Biotechnology(BIOTEC), National Science and Technology and Development Agency(NSTDA), Paholyothin Rd., Klong 1, Klong Luang, Pathumthani 12120, Thailand

Received 17 March 2003, Accepted 4 June 2003

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 ed 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 trans of apisimin, and one and two partial trans of α-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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