title: Expression of recombinant AccMRJP1 protein from royal jelly of Chinese honeybee in Pichia pastoris and its proliferation activity in an insect cell line.
abstract: 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.
authors: Department of Food Science and Nutrition, Zhejiang University 268 Kaixuan Road, Hangzhou, Zhejiang, China 310029. shenlirong@zju.edu.cn
authors: Lirong
authors: L
authors: Shen
authors: Weiguang
authors: W
authors: Zhang
authors: Feng
authors: F
authors: Jin
authors: Liwen
authors: L
authors: Zhang
authors: Zhengxian
authors: Z
authors: Chen
authors: Liang
authors: L
authors: Liu
authors: Laurence D
authors: LD
authors: Parnell
authors: Chao-Qiang
authors: CQ
authors: Lai
authors: Duo
authors: D
authors: Li
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D001516:Bees / Q000737:chemistry*; D002460:Cell Line; D049109:Cell Proliferation; D017931:DNA Primers; D018076:DNA, Complementary / Q000235:genetics; D005227:Fatty Acids / Q000737:chemistry*; D006023:Glycoproteins / Q000235:genetics / Q000502:physiology*; D019476:Insect Proteins / Q000235:genetics / Q000502:physiology*; D007313:Insecta; D008969:Molecular Sequence Data; D010843:Pichia / Q000235:genetics*; D016133:Polymerase Chain Reaction; D000099219:Royal Jelly
chemical_list: D017931:DNA Primers; D018076:DNA, Complementary; D005227:Fatty Acids; D006023:Glycoproteins; D019476:Insect Proteins; D000099219:Royal Jelly; C559157:MRJP1 protein, Apis mellifera