Sequence analysis of functional Apisimin-2 cDNA from royal jelly of Chinese honeybee and its expression in Escherichia coli
Apisimin is one of the functional peptides from royal jelly. The aim of this study was to analyze and in vitro express a new gene encoding Acc-apisimin-2 from Chinese honeybee (Apis cerana cerana) in Eschertchia coli.Ninety-six clones containing apisimin expressed sequence tag (EST) were identified from 8568 effective ESTs of the cDNA library of Chinese honeybee worker heads. The coding region of the matured peptide from one clone containing Acc-apisimin-2 gene was sub-cloned into the prokaryotic expression vector pGEX-4T-2. The recombinant vector then was transformed into E. coli BL21 (DE3) for expression. The expression product was analyzed with SDS-PAGE and Western blot. The total length of the Acc-apisimin-2 cDNA was 379 bp, containing an open-reading frame (ORF) of 237 nucleotides encoding a 78 amino acid residue precursor. The Acc-apisimin-2 gene shared 100％ homologies with Am-apisimin from A. mellifera, but 93％ and 91％ homologies with Aciapisimin from A. cerana indica and the previously reported Acc-apisimin-1 sequence (AY278991) on a nucleotide level, respectively. The GST-Acc-apisimin-2 fusion protein expressed in the recombinant vector was about 31 kDa in size and accumulated up to about 22.1％ of the total bacterial proteins. About 50％ of the recombinant protein was soluble. The fusion protein purified through affinity chromatography was cross reactive with GST antibody, which confirmed the successful expression of GST-Acc-apisimin-2.
Apis cerana cerana royal jelly Acc-apisimin-2 sequence analysis prokaryotic expression
Lirong Shen Liping Xing Yuxiao Yang Qikang Gao
Department of Food Science and Nutrition, Zhejiang University, China Institute of Insect Sciences, Zhejiang University, China
国际会议
国际营养科学联盟第八届临床营养学大会暨第五届亚太临床营养学会大会
杭州
英文
222-226
2006-10-01（万方平台首次上网日期，不代表论文的发表时间）