Schmidt U, Kenklies J, Rudolph R, Böhm G
Institut für Biotechnologie, Martin-Luther-Universität Halle-Wittenberg, Halle (Saale), Germany.
Biol Chem. 1999 Mar;380(3):397-401. doi: 10.1515/BC.1999.053.
For the development of gene therapy protocols based on polyomavirus-like particles, we describe a method for fluorescence labelling of virions in order to study virus-cell interactions preceding gene delivery. Site-specific fluorescence labelling of polyomavirus-like particles is achieved via a single cysteine residue and maleimide conjugates of fluorescence dyes (fluorescein, Texas Red). Polyomavirus-like particles can be assembled in vitro from recombinant capsomers produced in E. coli. Since the assembly process is independent of disulfide bond formation, all cysteine residues of the wild-type protein were replaced by serines, and a new unique cysteine residue was introduced for the attachment of the fluorescence marker.
为了开发基于多瘤病毒样颗粒的基因治疗方案，我们描述了一种用于病毒粒子荧光标记的方法，以研究基因传递之前的病毒-细胞相互作用。多瘤病毒样颗粒的位点特异性荧光标记是通过荧光染料（荧光素、德克萨斯红）的单个半胱氨酸残基和马来酰亚胺缀合物实现的。多瘤病毒样颗粒可以从大肠杆菌中产生的重组衣壳蛋白体外组装而成。由于组装过程不依赖于二硫键的形成，野生型蛋白的所有半胱氨酸残基都被丝氨酸取代，并引入了一个新的独特半胱氨酸残基用于连接荧光标记。
