M13 is a filamentous bacteriophage with a 6 nm thick and 1 µm length tubular structure. Approximately, 3000 subunits of PVIII, the major coat protein form a cylinder that encapsulates the M13 single-stranded DNA genome. Among capsid proteins sequences, PVIII gene is particularly amenable for the expression of recombinant peptides on the phage surface. It has been identified that thiol rich peptides are able to interact with heavy metals. In this work, we incorporated a heavy metal binding peptide to PVIII producing a viral particle capable of interacting with cadmium. For this purpose, we designed a chimeric gene composed of the coding sequence of a cysteine rich peptide linked to PVIII and cloned into a phagemid vector. Then, we established the optimal conditions for phage production by regulating the expression of the chimeric PVIII protein by using an isopropyl β-D-1-thiogalactopyranoside (IPTG) inducible system. The cysteine rich recombinant phage interacted with cadmium in a highly efficient manner. Considering this novel property, the recombinant phage produced in this work could be used as a nanotechnology tool in the development of affinity columns, concentration systems for cadmium analysis or bioremediation.

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通过在PVIII上展示富含半胱氨酸的肽生成结合镉的丝状噬菌体

M13是一种丝状噬菌体，具有6 nm厚和1 µm长的管状结构。PVIII的大约3000个亚基（主要外壳蛋白）形成了一个圆柱体，该圆柱体封装了M13单链DNA基因组。在衣壳蛋白序列中，PVIII基因特别适合在噬菌体表面表达重组肽。已经确定，富含硫醇的肽能够与重金属相互作用。在这项工作中，我们将重金属结合肽结合到PVIII上，从而产生能够与镉相互作用的病毒颗粒。为此，我们设计了一个嵌合基因，该基因由与PVIII连接并克隆到噬菌粒载体中的富含半胱氨酸的肽的编码序列组成。然后，我们通过使用异丙基β-D-1-硫代半乳糖吡喃糖苷（IPTG）诱导系统调节嵌合PVIII蛋白的表达，建立了噬菌体生产的最佳条件。富含半胱氨酸的重组噬菌体以高效方式与镉相互作用。考虑到这种新特性，这项工作中产生的重组噬菌体可以作为纳米技术工具用于亲和柱，镉分析或生物修复浓缩系统的开发中。