Fil'chikov M V, Osmakov D I, Logovskaia L V, Sykilinda N N, Kadykov V A, Kurochkina L P, Mesianzhinov V V, Bernal R A, Miroshnikov K A
Bioorg Khim. 2009 Nov-Dec;35(6):808-15.
The virulent P. aeruginosa bacteriophage SN belongs to the PB1-like species of the Myoviridae family. The comparatively small (66391 bp) DNA genome of this phage encodes 89 predicted open reading frames and the proteome involves more than 20 structural proteins. A 3D model of the phage capsid to approximately 18 A resolution reveals certain peculiarities of capsomer structure typical of only this bacteriophage species. In the present work recombinant structural proteins SN gp22 and gp29 were expressed and purified; and specific polyclonal antibodies were obtained. Immune-electron microscopy of purified phage SN using secondary gold-conjugated antibodies has revealed that gp29 forms a phage sheath, and gp22 decorates the capsid. Precise identification of multicopy major capsid proteins is essential for subsequent construction of gene-engineered phages bearing non-native peptides on their surfaces (phage display).
强毒铜绿假单胞菌噬菌体SN属于肌尾噬菌体科PB1样种。该噬菌体相对较小(66391 bp)的DNA基因组编码89个预测的开放阅读框,其蛋白质组涉及20多种结构蛋白。噬菌体衣壳的三维模型分辨率约为18 Å,揭示了仅该噬菌体物种特有的衣壳粒结构的某些特性。在本研究中,表达并纯化了重组结构蛋白SN gp22和gp29;并获得了特异性多克隆抗体。使用二次金标抗体对纯化的噬菌体SN进行免疫电子显微镜观察发现,gp29形成噬菌体尾鞘,而gp22装饰衣壳。准确鉴定多拷贝主要衣壳蛋白对于随后构建表面带有非天然肽的基因工程噬菌体(噬菌体展示)至关重要。