人源性exCAR原核表达蛋白体外阻断腺病毒感染实验研究

In vitro experimental study of blocking effect on adenovirus infection of prokaryotic expressed protein of human exCAR

目的在培养乳鼠心肌细胞上检测人源性柯萨奇病毒-腺病毒受体胞外区(exCAR)大肠杆菌原核表达蛋白阻断腺病毒感染的活性。方法纯化人源性exCAR表达蛋白并通过透析复性,进一步体外实验,在腺病毒感染乳鼠心肌细胞模型上检测表达蛋白阻断腺病毒感染效率。结果获得纯化表达蛋白exCAR,经复性后,在体外实验中检测到其阻断腺病毒感染心肌细胞效应,其终浓度为500ng/mL时,获得最高阻断效率约75%。结论本研究获得了具有生物活性的exCAR表达蛋白,为腺病毒感染防治提供了新的思路和基础。

Objective To test the blocking effect on adenovirus infection of the expressed protein of extra-cellular region of human Coxsackie-adenovirus receptor in neonatal rat cardiac myocytes. Methods The human exCAR protein was expressed in E. coli expression system pET-28a（ ＋）/BL21, and the expression protein was purified and renatured by dialysis, and then the blocking effect on adenovirus infection to neonatal rat cardiac myocytes was measured. Results The purify expressed recombinant exCAR protein was obtained and renatured,and it＇s blocking effect on adenovirus infection was identified on neonatal rat cardiac myocytes. The result showed that as the final concentration of exCAR was 500ng/mL,it had the most marked blocking effect,which was about 750%. Conclusion The exCAR recombinant protein with bio-aetive is obtained,and this work provides a basis and materials for a new therapy way of the adenovirus infection diseases.