摘要
研究以AFB1-BSA免疫的小鼠脾脏细胞为试验材料,利用RT-PCR技术,克隆了抗AFB1抗体重链和轻链可变区基因VH和VL,利用连接肽(Gly4Ser)3将VH和VL链接成单链抗体基因scFv,通过噬菌体展示载体pCANTAB-5E携带将其电转化E.coli TG1,经氨苄青霉素平板筛选,构建了库容为2.13×109 cfu/μg DNA的噬菌体单链抗体库,抗体库的克隆阳性率达到100%,且多样性良好,为高活性抗AFB1单链抗体的筛选提供了材料基础。
The immunitied spleen cells of mouse by AFB1-BSA were used for test materials,resisted AFB1 antibody heavy chain VH and light chain gene VL were cloned by using RT-PCR,the linker peptide（Gly4Ser）3 was used to link VH and VL into a single chain gene scFv,through phage vectors carrying the pCANTAB-5E transformated into E.coli TG1 by electricity,adopted ampicillin flat screening,contenting 2.13×109 cfu/μg DNA of phage single-chain antibody libraries were constructed,the positive clone of antibody library reached 100%,provided the materials base for screening of highly active anti-AFB1 antibody.
出处
《齐齐哈尔大学学报（自然科学版）》
2010年第6期76-80,共5页
Journal of Qiqihar University(Natural Science Edition)
基金
黑龙江省教育厅项目(1151hz025)
