Soluble Expression and Purification of Q۵۹L Mutant L-asparaginase in the Presence of Chaperones in SHuffle™ T۷ strain
- سال انتشار: 1400
- محل انتشار: مجله بین المللی آزمایشگاه پزشکی، دوره: 8، شماره: 2
- کد COI اختصاصی: JR_JIML-8-2_007
- زبان مقاله: انگلیسی
- تعداد مشاهده: 242
نویسندگان
Department of Medical Biotechnology, Faculty of Medicine, Shahid Sadoughi University of Medical Sciences, Yazd, Iran
Department of Medical Biotechnology, Faculty of Medicine, Shahid Sadoughi University of Medical Sciences, Yazd, Iran
Department of Medical Biotechnology, Faculty of Medicine, Shahid Sadoughi University of Medical Sciences, Yazd, Iran
چکیده
Background and Aims: Q۵۹L mutant of L-asparaginase enzyme from Escherichia coli (E. coli) has been introduced with lower side effects. This version of the enzyme might have potential applications in the treatment of leukemia patients. We utilized SHuffle T۷ strain of E. coli, to produce the mutant enzyme in the presence of chaperone molecules. Materials and Methods: Q۵۹LAsp gene was cloned into pET۲۸a expression vector, and two strains of E. coli (BL۲۱ DE۳ and SHuffle T۷ strains) were used to produce recombinant protein. In parallel, PG-Tf۲ plasmid was cloned into the same strains, and the effect of trigger factor chaperone and groELS chaperonines was studied. The his-tagged recombinant protein was expressed and purified using nickel affinity chromatography. The amount of recombinant protein which is produced in each condition was determined and compared. Results: The amount of soluble recombinant protein was enhanced in the presence of chaperones in both strains of E. coli. SHuffle T۷ strain produced more soluble recombinant protein in the soluble state than BL۲۱ DE۳ strain. So the best condition for the production of soluble recombinant Q۵۹L mutant protein was the use of PG-Tf۲ plasmid in the SHuffle T۷ strain. Conclusion: Application of the new strain SHuffle T۷, with chaperones simultaneously showed better results in the production of Q۵۹L mutant version of L-Asparaginase.کلیدواژه ها
Asparaginase, Chaperone, E. coli, Shuffle, آسپارژیناز, لوسمی, چاپرون, سویه shuffleاطلاعات بیشتر در مورد COI
COI مخفف عبارت CIVILICA Object Identifier به معنی شناسه سیویلیکا برای اسناد است. COI کدی است که مطابق محل انتشار، به مقالات کنفرانسها و ژورنالهای داخل کشور به هنگام نمایه سازی بر روی پایگاه استنادی سیویلیکا اختصاص می یابد.
کد COI به مفهوم کد ملی اسناد نمایه شده در سیویلیکا است و کدی یکتا و ثابت است و به همین دلیل همواره قابلیت استناد و پیگیری دارد.