南京大学学报(自然科学版) ›› 2015, Vol. 51 ›› Issue (5): 10841090.
段宁骏1,2,吕万胜1,2,贡佳1,郑伟娟1,2*,华子春1,2*
Duan Ningjun1,2LyuWansheng1,2Gong Jia1Zheng Weijuan1,2*Hua Zichun1,2*
摘要: 透明质酸是一种长链大分子糖胺聚糖,存在于各组织器官中,是细胞外基质的重要组成部分。透明质酸不仅能够与其他胞外多糖及蛋白构成骨架,参与组织器官的结构形成,也可以与细胞表面相关蛋白受体相互作用,通过相关信号通路影响细胞的生存、增殖以及迁移。研究证实,透明质酸与肿瘤细胞的生存、迁移,肿瘤血管发生等均有密切关系,组织中透明质酸含量的变化也与某些病理状况如慢性炎症相关。因此,细胞表面或组织中的透明质酸含量的定量检测,对于疾病的诊断及预后都具有重要意义。目前透明质酸的检测方法主要依靠抗体,通过酶联免疫检测体液中的透明质酸,而缺乏在细胞或组织水平上检测透明质酸的手段。本研究将三种人源的透明质酸结合蛋白(P32,CDC37以及RHAMM)与绿色荧光蛋白(EGFP)进行融合,在大肠杆菌中进行可溶性表达,经亲和纯化,获得了三种能够与透明质酸结合的蛋白探针。通过在溶液与细胞水平上比较与透明质酸结合的能力,从中筛选出了最适合用来进行透明质酸检测的荧光蛋白探针RHC-EGFP。
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