摘要
目的:构建日本血吸虫单克隆抗独特型抗体NP30重链可变区6×CDR3基因/IL-2融合基因,以观察其对动物的抗血吸虫感染保护作用。方法:通过PCR方法体外扩增NP30 6×V_HCDR3和IL-2基因,经测序后先后重组入原核表达质粒pET-28a(+)相应的位点上,将构建正确的6×V_HCDR3/IL-2-pET-28a(+)表达质粒转化E.coli BL21,IPTG诱导表达。结果:构建的6×V_HCDR3/IL-2融合基因中,6×V_HCDR3和IL-2序列均正确,融合基因经IPTG诱导表达重组蛋白,十二烷基磺酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE)分析分子质量约为30ku,表达量约占菌体总蛋白的20％。结论:成功构建并原核表达了NP30 6×V_HCDR3/IL-2融合基因。
Objective: To construct and express the fused gene of NP30 6 × VHCDR3 and IL-2 for observing the protective immunity in mice induced by the fusion protein. Methods: The NP30 6 × VHCDR3 and IL-2 gene fragments were amplified by PCR and inserted into corresponding sites of expression vector pET-28a( + ), and the fusion gene was induced to express in E. coli BL21. Results: The sequences of NP30 6 × VHCDRS and IL-2 were correct in the fusion gene. The fusion protein expressed in E. coli amounted to 20% of the total bacterial protein and it was displayed as a single band of Mr 30 ku by SDS-PAGE. Conclusion: The fusion gene of NP30 6× VHCDR3/IL-2 was constructed and expressed successfully.
出处
《南京医科大学学报（自然科学版）》
CAS
CSCD
北大核心
2003年第4期299-301,共3页
Journal of Nanjing Medical University(Natural Sciences)
基金
国家自然科学基金资助项目(39970670)
