摘要
目的 :获得抗黏性放线菌Ⅰ型菌毛单抗特异性单链抗体。方法 :从抗黏性放线菌Ⅰ型菌毛蛋白杂交瘤细胞系 (mABCM)中分离总RNA ,经RT -PCR体外扩增重、轻链可变区基因 ,通过一连接肽拼接形成ScFv基因 ,经菌毛蛋白抗原亲和筛选出特异性阳性克隆进行核苷酸序列测定及氨基酸序列推导。结果 :所构建的单链抗体基因为拼接正确的VH-Linker-VL 结构 ,无错义及移码读框 ,且含有编码 (Gly4Ser) 3的连接肽基因及维持抗体结构所必需的半胱氨酸残基。结论 :所构建的单链抗体为VH-Linker-VL 结构 ,含有连接肽基因和半胱氨酸残基 ,具有单链抗体特征。
AIM:To construct single chain antibody fragment from mAb CM against type Ⅰ fimbriae of actinomyces viscosus.METHODS:Reverse transcription and polymerize chain reaction were used to clone the V region genes of mAb CM(V H,V L).The cloned V H and V L were constructed into a single chain antibody(ScFv).Nucleotide sequence of the constructed ScFv was sequenced and its amino acid was induced.RESULTS:The construction of the single chain antibody fragment was V H-Linker-V L.CONCLUSION:Single chain antibody fragment of mAb CM was successfully constructed. [
出处
《牙体牙髓牙周病学杂志》
CAS
2003年第10期548-551,共4页
Chinese Journal of Conservative Dentistry
基金
本课题由上海市教育委员会曙光计划 99SG41资助
关键词
单链抗体
噬菌体抗体库
黏性放线菌
I型菌毛蛋白
single chain Fv fragment(ScFv)
phage antibody library
actinomyces viscosus
type Ⅰfimbriae
