Cloning of human survivin gene into pET۲۸a bacterial vector
- سال انتشار: 1399
- محل انتشار: چهارمین کنگره بین المللی و شانزدهمین کنگره ملی ژنتیک
- کد COI اختصاصی: CIGS16_389
- زبان مقاله: انگلیسی
- تعداد مشاهده: 428
نویسندگان
Department of Biochemistry, Faculty of Biological Sciences, Tarbiat Modares University, Tehran, Iran
Department of Biochemistry, Faculty of Biological Sciences, Tarbiat Modares University, Tehran, Iran
Department of Biochemistry, Faculty of Biological Sciences, Tarbiat Modares University, Tehran, Iran
چکیده
Background and Aim: Apoptosis is a form of programmed cell death leading to the elimination of cell without releasing harmful substances into the surrounding area. Inhibitor of apoptosis are a family of proteins that mainly block programmed cell death. The human IAP family consist of ۸ members. Survivin is a smallest well-known inhibitor of apoptosis proteins family member. Survivin overexpression occurs in many different cancer types but not in the normal tissue except embryonic tissue. Survivin may be used as a new marker to stratify cancerous patients for more optimal treatment modalities, or can be a promising new target for therapy. In this research, we cloned human survivin gene into pET۲۸a bacterial vector.Methods: Specific forward and reverse primers were designed. PCR was performed and survivin gene amplified by specific primers and tempelet. PCR product of survivin and pET۲۸a vector were digested by HindIII/NheI restriction enzymes and survivin was ligated into the HindIII/NheI digested/dephosphorylated pET۲۸a vector. Then, the ligation product was transformed into the E.coli DH۵a competent cells and screened by antibiotic selection marker (kanamycine).Results: Positive colonies were selected by colony PCR and screened by double digestion of isolated plasmid. One positive colony was sequenced and confirmed the inserted DNA.Conclusion: Human survivin gene was cloned in pET۲۸a vector. After validation by sequencing, pET۲۸a/survivin product was transformed into the E.coli BL۲۱(DE۳) competent cells to study recombinant protein expression.کلیدواژه ها
Survivin, Cloning, pET۲۸a.مقالات مرتبط جدید
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