摘要
目的从噬菌体构象型7肽库中筛选人HMGB1-Bbox的抑制性小肽。方法以重组人HMGB1-Bbox为靶分子对噬菌体构象型7肽库进行6轮亲和筛选,获得Bbox结合的克隆,并经ELISA验证。选取亲和力高的克隆进行DNA测序,并推导出呈现的多肽序列,通过IL-6 ELISA检测噬菌体呈现的小肽对人HMGB1-B box致炎功能的抑制作用。结果经过6轮亲和筛选,噬菌体的回收率增加,阳性克隆得到富集。挑选15个结合力强的克隆进行测序,推导出2个多肽序列。所获两个阳性噬菌体克隆能特异性地抑制人HMGB1-B box刺激THP-1细胞产生炎症因子的能力。结论获得了噬菌体呈现的能够抑制人HMGB1-B box的两个小肽。
Objective To screen and characterize antagonistic peptides inhibiting hHMGB1-B box. Methods C7C phage display peptide library was screened with hHMGB1-B box. Positive phage clones were identified by ELISA. Results After 6 rounds of screening, the ratio of output to input and specific enrichment was gradually increased and positive clones were enriched. Two phage clones inhibiting the activity of HMGB1-B box were identified. Conclusion Two phage display antagonist peptides blocking HMGB1-B box were obtained.
出处
《医学分子生物学杂志》
CAS
CSCD
2010年第1期11-15,共5页
Journal of Medical Molecular Biology
基金
国家自然科学基金(No.30670411),军队“十一五”医药卫生研究基金资助项目(No.06H027),重庆市自然科学基金(No.CSCT2008BB5022)