Cloning of human high-temperature requirement A2 (hu-HtrA2) gene in a prokaryotic host

سال انتشار: 1397
نوع سند: مقاله کنفرانسی
زبان: انگلیسی
مشاهده: 308

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شناسه ملی سند علمی:

BIOCONF20_230

تاریخ نمایه سازی: 28 اردیبهشت 1398

چکیده مقاله:

HtrA2/omi is a mitochondrial serine protease which is induced by the stress and encoded by the nucleus. This protease protects the cell survival by maintaining the mitochondrial homeostasis. The HtrA2/omi has a role in cell death caused by apoptosis, necrosis, and anoikis. Disordered HtrA2/omi appears in neurological diseases, cancer and arthritis. For this reason, studying of the structure and function of this enzyme can be a way of preventing and treating diseases and disorders caused by this enzyme. In this study, the HtrA2/omi gene was cloned in pET21a (+) (expression vector). For cloning into pET21a (+) one should clone a target gene between the NdeI (forward primer) and the XhoI (reverse primer) sites in pET21a (+). After PCR, the PCR product was digested by NdeI and XhoI restriction enzymes and then ligated to the vector. Finally, E.coli TOP 10 strain cells were transformed by the ligation product and incubated at 37°C for 16h. After observation of colonies, some colonies were randomly selected and prepared for colony PCR. The results of colony PCR have confirmed the cloning of HtrA2/omi in pET21a (+). Also, the recombinant vectors were extracted for sequencing. The expression and purification of the recombinant protein is underway.

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نویسندگان

Samira Shaterian,

Department of Biology, College of Fundamental Science, Azarbaijan Shahid Madani University, Tabriz, Iran

Saeed Najavand

Department of Biology, College of Fundamental Science, Azarbaijan Shahid Madani University, Tabriz, Iran

Mohammad Pazhang

Department of Biology, College of Fundamental Science, Azarbaijan Shahid Madani University, Tabriz, Iran