Expression of Recombinant AccMRJP1 Protein from Royal Jelly of Chinese Honeybee in Pichia pastoris and Its Proliferation Activity in an Insect Cell Line
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- 作者：Lirong Shen ; Weiguang Zhang ; Feng Jin ; Liwen Zhang ; Zhengxian Chen ; Liang Liu ; Laurence D. Parnell ; Chao-Qiang Lai ; Duo Li
- 刊名：Journal of Agricultural and Food Chemistry
- 出版年：2010
- 出版时间：August 25, 2010
- 年：2010
- 卷：58
- 期：16
- 页码：9190-9197
- 全文大小：1004K
- 年卷期：v.58,no.16(August 25, 2010)
- ISSN：1520-5118
文摘
Major royal jelly protein 1 (MRJP1) is the most abundant member of the major royal jelly protein (MRJP) family of honeybee. Mature MRJP1 cDNA of the Chinese honeybee (Apis cerana cerana MRJP1, or AccMRJP1) was expressed in Pichia pastoris. SDS-PAGE showed that recombinant AccMRJP1 was identical in molecular weight to the glycosylated AmMRJP1 from the Western honeybee (Apis mellifera). Western blots probed with anti-AccMRJP1 antibody demonstrated that recombinant AccMRJP1 and soluble protein of the Western honeybee RJ (AmSPRJ) contained immunoreactive MRJP1. The 57 kDa protein in AmSPRJ contained an N-terminal amino sequence of N-I-L-R-G-E, which is identical to that previously characterized in AmMRJP1. The molecular weight of recombinant AccMRJP1 was decreased from 57 to 48 kDa after deglycosylation, indicating that AccMRJP1 was glycosylated. The recombinant AccMRJP1 significantly stimulated Tn-5B-4 cell growth, similar to AmSPRJ and fetal bovine serum, and affected cell shape and adhesion to the substrate.