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Abstract
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HtrA2/omi is the mitochondrial serine protease, induced by the stress and encoded by the nucleus. It protects the cell survival by maintaining the mitochondrial homeostasis. The HtrA2/omi has a role in cell death by apoptosis, necrosis, and anoikis. Disordered HtrA2/omi has appeared in neurological diseases, cancer and arthritis. so, The study of the structure and function of the enzyme can be an important and effective step in preventing and treatment of diseases caused by this enzyme. In this study, The coloning of HtrA2/omi gene in pET21a(+) (expression vector) has been done. For the gene coloning, coloning primers were designed with site to NdeI (forward primer) and XhoI (reverse primer). After PCR, The PCR product was digested by NdeI and XhoI restriction enzymes and then, the digested fragments were extracted from the agarose gel and then the ligation was performed. E.coli TOP 10 strain cells were transformed by ligated product and incubated in 37°C for 16h. After observation of colonies, some colonies were randomly selected and prepared for colony PCR. The results of colony PCR confirmed the coloning of HtrA2/omi in pET21a(+). Also, the recombinant vectors were extracted for sequencing. The expression and purification of this gene is underway.
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