摘要
目的研究大肠杆菌DNA片段对HBsAg的免疫刺激作用。方法将大肠杆菌DNA片段,通过PCR补齐、加尾后,连于pGEMT载体,再进行PCR扩增后得到100～250bp的dsDNA片段,将扩增出的片段(100μg)与HBsAg混合免疫ICR小鼠。于免后8周检测其抗体水平,选择抗体水平最高的组进行重复实验,对其中最高的一组测序,并分析其序列特征。结果在235个DNA片段中,有的片段免疫刺激作用显著高于铝佐剂和CpGODN1826对照组。该DNA片段除含有1个符合免疫刺激模式的基序外,其余12个CpG二核苷酸均为非回文结构。结论大肠杆菌DNA中含CpGODN的片段对HBsAg具有免疫刺激作用。
Objective To study the immunostimulating effect of E.coli DNA fragments on HBsAg.Methods Extract DNA fragment from E.coli,amplify by PCR and insert into vector pGEM-T for the amplification of dsDNA fragments at lengths of 100-250 bp by PCR.Mix the amplified dsDNA fragments (100 μg) with HBsAg to immunize ICR mice,using aluminium adjuvant and CpG ODN1826 as controls.Determine the antibody level 8 weeks after immunization.Retest the fragments inducing the highest antibody level.Sequence the fragment inducing the highest antibody level in retest and analyze the characteristic of its sequence.Results The antibody levels induced by a part of amplified DNA fragments were significantly higher than those induced by HBsAg+aluminium adjuvant and HBsAg+CpG ODN1826.Except an active CpG motif,No.187 fragment inducing the highest antibody level in retest contained 12 nonpalindromic CpG motifs.Conclusion The E.coli DNA fragment containing CpG ODN showed immunostimulating effect on HBsAg.
出处
《中国生物制品学杂志》
CAS
CSCD
2005年第3期225-226,240,共3页
Chinese Journal of Biologicals
