摘要
利用不同拓扑学结构的质粒 ,分别转化 E.coli感受态细胞、转染 SK6 - 6细胞和免疫小鼠 ,比较了不同拓扑学结构质粒对转化效率、转染率和对免疫应答水平的影响。结果显示 ,超螺旋占 5 0 %比例的猪瘟病毒 E2基因表达质粒p CDSW转化 E.coli感受态细胞的效率比开环的 p CDSW高 8倍 ,超螺旋比例占 80 %的 p VAXE2 IL- 2质粒比酶切线性化质粒转化率高 2 2倍 ;以荧光蛋白基因为报告基因 ,超螺旋比例占 80 %的荧光蛋白真核表达质粒转染真核细胞的效率是线性化质粒的 12倍 ;以猪瘟病毒主要保护性抗原 E2基因表达质粒 ,分别制备成超螺旋和开环形式占优势的质粒 ,免疫昆明鼠 ,抗体检测结果显示 。
The transformation/transfection efficiency of different topology plasmids for E.coli competence cell/SK cell line indicated the supercoiled plasmid DNA(closed circular DNA,cccDNA) was higher than that of the nicked DNA and linearieed DNA.The fact that the proportion of supercoiled plasmid was closely related to preparation method was also demonstrated in this study.By detecting the humoral immune response against CSFV in mice induced by the same plasmid with different topology(prepared with different methods) administered intramuscularly,it was found that the induced antibody level was positively correlated with the proportion of supercoiled plasmid.This result indicated that the topology of plasmid exerted a significant influence on DNA-mediated humoral immune response.
出处
《中国兽医学报》
CAS
CSCD
北大核心
2004年第3期250-251,共2页
Chinese Journal of Veterinary Science
基金
国家"8 63"计划项目 ( 2 0 0 1AA2 13 14 1)
