Department of Microbiology, Islamic Azad University, Jahrom, Iran.
Department of Biology, Islamic Azad University, Damghan, Iran.
Prep Biochem Biotechnol. 2021;51(1):9-15. doi: 10.1080/10826068.2020.1762215. Epub 2020 May 12.
Cancer is considered as a disease with high rates of mortality and morbidity. The limitations and side effects of common treatments have prompted the need for innovative cancer therapies. Furthermore, selectivity and targeting of cancer cells are crucial factors to successful treatment of cancer. One of these methods is the use of bacterial toxins including toxin to aid cancer therapy. This toxin is composed of three polypeptides: protective factor (PA), lethal factor (LF), and edema factor (EF). PA can bind to various surface receptors of all types of human cells and it internalizes the lethal factor and edema factor subunits of the toxin in the cytosol. In the present study, we cloned and expressed the lef gene of as the lethal part of the toxin in WB600 by a shuttle expression vector PHT4. The rLF made in is efficiently secreted by the host into the culture medium which facilitates downstream processing. The rLF can be used to study cancer treatment. EF: edema factor; LF: lethal factor; PA: protective factor; rLF: recombinant lethal factor; rPAm: recombinant protective factor mutants; uPA: urokinase-type plasminogen activator; uPAR: urokinase-type plasminogen activator receptor.
癌症被认为是一种死亡率和发病率都很高的疾病。常见治疗方法的局限性和副作用促使人们需要创新的癌症治疗方法。此外，癌症细胞的选择性和靶向性是癌症治疗成功的关键因素。其中一种方法是使用细菌毒素，包括志贺毒素来辅助癌症治疗。这种毒素由三个多肽组成：保护因子（PA）、致死因子（LF）和水肿因子（EF）。PA 可以与各种人类细胞的表面受体结合，并将毒素的致死因子和水肿因子亚单位内化到细胞质中。在本研究中，我们通过穿梭表达载体 PHT4 将志贺氏菌的 lef 基因克隆并表达为毒素的致死部分，在 WB600 中。由宿主高效分泌到培养基中的 rLF 有利于下游加工。rLF 可用于研究癌症治疗。EF：水肿因子；LF：致死因子；PA：保护因子；rLF：重组致死因子；rPAm：重组保护因子突变体；uPA：尿激酶型纤溶酶原激活物；uPAR：尿激酶型纤溶酶原激活物受体。
