[Studies on protein-based identification method of genetically modified capsicum].
The detection system based on protein is a method to evaluate the safety of genetically modified foods (GMF). Using cecropin BD gene in capsicum, a detecting method was set up. It is a system of evaluating the real expressive condition and safety of the foreign target protein of GMF. In this studies, with the preformative technic method, a satisfactory results by making use of hemolymph of immunized pupae of Antheraea pernyi as standard experimental material was achieved, comparing with the realities of the goal protein expressive condition of cecropin D gene in capsicum. The detecting steps were as following: the goal protein from material was extracted roughly, then with CM-Sepharose-FF ion-exchange chromatography twice, the goal protein was purified moderately. The purified product was identified by detecting the anti-bacterial activity, electrophoresis, biological auto-photography of the goal protein and MADDI-TOF mass spectrum. The results showed that the expressive foreign target protein in transgenic capsicum was in accordance with standard protein in the physical and chemical property, anti-bacterial activity and molecular weight. It indicated that expression of the target gene in capsicum is real, it corresponded to expected value. The separation, purification and identification methods of cecropin D were established in the study. By means of the comparative experiments about anti-bacterial activity and molecular weight of anti-bacterial peptide(ABP) from GM-capsicum and hemolymph of immunized pupae of Antheraea pernyi, the identification method of target protein from GM-capsicum was set up. The method is easy to be operated, fast and feasible.
Liu
Jianjun
J
Shenzhen Disease Prevention and Control Center, Shenzhen 518020, China.
Deng
Pingjian
P
Fang
Shisong
S
Zhao
Jin
J
D002212:Capsicum / Q000737:chemistry* / Q000235:genetics; D030841:Food, Genetically Modified / Q000009:adverse effects; D019476:Insect Proteins / Q000235:genetics; D014674:Plant Proteins, Dietary / Q000009:adverse effects / Q000032:analysis*; D030821:Plants, Genetically Modified / Q000737:chemistry*
D019476:Insect Proteins; D014674:Plant Proteins, Dietary; C036149:cecropin D protein, Hyalophora cecropia