摘要
目的:对天花粉蛋白(Trichosanthin,TCS)进行定点突变,比较其与野生型TCS(wTCS)间的生物活性及免疫原性差异。方法:选择TCS分子上可能的抗原决定簇位点(YFF81-83和KR173-174)进行定点突变,并将所构建的突变体(TCSYFF-KR)在大肠杆菌中表达及纯化。随后将其与wTCS比较,分析其DNA酶活性、致核糖体失活活性和免疫原性。结果:构建的突变体TCSYFF-KR其活性与wTCS活性几乎相当,而免疫原性有所降低。结论:通过该方法改造TCS是可行的,所构建的突变体TCS值得进一步研究。
Objective： To construct a trichosanthin （TCS） mutant and analyze its bioactivities and immunogenicity. Methods： By computer modeling, two sites （YFF81 -83 and KR173- 174） were identified as potential antigenic determinant of TCS. A TCS mutant named for TCSYFF-KR was constructed by site - directed mutagenesis and expressed in E. coli BL21 （DE3）. The bioactivities and immunogenicity of the TCS mutant were detected. Results： The bioactivities of the TCS mutant were similar to wild - type TCS （ wTCS）, but with much lower immunogenicity . Conclusion： The constructed TCS mutant should be further studied.
出处
《西北国防医学杂志》
CAS
2008年第4期254-256,共3页
Medical Journal of National Defending Forces in Northwest China
基金
陕西省科技计划资助项目(2003K10G9)
关键词
天花粉蛋白
定点突变
生物活性
免疫原性
Trichosanthin
Site - directed mutagenesis
Bioactivity
Immunogenicity
