关键词: 湿疹, 单纯疱疹病毒属, 包膜糖蛋白

Abstract: Zhang Yu*, Yao Xu, Gu Hanyan, Wang Baoxi, Liu Jun. *Institute of Dermatology, Chinese Academy of Medical Sciences and Peking Union Medical College, Nanjing 210042, China Corresponding authors: Yao Xu, Email: dryao_xu@126.com; Liu Jun, Email: drliu_jun@126.com 【Abstract】 Objective To synthesize herpes simplex virus （HSV） envelope glycoprotein gC using gene engineering techniques, and to verify its expression. Methods Two separate parts of the HSV envelope glycoprotein gC, i.e., GC-F and GC-R, were respectively synthesized. The GC-F and GC-R genes were synthesized, subcloned into the expression vectors pSumo-Mut （containing recognition sequences for endonucleases Stu1 and XhoI） and pCzn1 （containing recognition sequences for endonucleases NdeI and XhoI） respectively to form the recombinant plasmids pSumo-Mut-GC-F and pCzn1-GC-R. E. coli BL21 Arctic Express （DE3） cells were transformed with the two recombinant plasmids separately. Isopropyl thiogalactoside （IPTG） was used to induce the expression of target protein which was subsequently purified by nickel affinity chromatography. Finally, Western blot was performed to verify the reactivity of the synthesized protein with the sera of HSV-1-positive patients. Results Both GC-F and GC-R genes were synthesized by a total gene synthesis method. As sodium dodecyl sulfate polyacrylamide gel electropheresis （SDS-PAGE） showed, the fusion proteins were mainly distributed in the sediment layer. The purity of GC-F and GC-R proteins was over 80% after purification by affinity chromatography. Western blot showed that both of the proteins were reactive with anti-HSV-1 antibody-positive sera. Conclusions Fusion expression vectors have been constructed for the gC protein, and IPTG successfully induces its expression. Moreover, the resulting proteins could react with anti-HSV-1 antibody-positive sera, and may serve as an ideal experimental material for next functional study.

Key words: Eczema, Simplexvirus, Glycoprotein C