University of Tehran Press Progress in Biological Sciences 1016-1058 4 2 2014 12 01 Apoptotic protease-activating factor 1 (Apaf-1) as a liable gene for spontaneous mutations in vitro 261 273 52304 10.22059/pbs.2014.52304 EN Taha Azad Department of Biochemistry, Faculty of Biological Sciences, Tarbiat Modares University, Tehran, Iran Amin Tashakor Department of Biochemistry, Faculty of Biological Sciences, Tarbiat Modares University, Tehran, Iran Mina Ghahremani Department of Plant Biology, Faculty of Biological Sciences, Tarbiat Modares University, Tehran, Iran Roohullah Hemmati Department of Biochemistry, Faculty of Biological Sciences, Tarbiat Modares University, Tehran, Iran Farangis Ataei Department of Biochemistry, Faculty of Biological Sciences, Tarbiat Modares University, Tehran, Iran Saman Hosseinkhani Department of Biochemistry, Faculty of Biological Sciences, Tarbiat Modares University, Tehran, Iran Journal Article 2014 09 10 The apoptotic protease-activating factor 1 (Apaf-1) receives the death signal in the intrinsic or<br />mitochondrial pathway of apoptosis. Upon the releasing of cytochrome c from the<br />intermembrane space of mitochondria and binding to Apaf-1 molecules, a heptameric<br />apoptosome complex is formed and triggers the downstream cascade of caspases. Here, for the<br />first time we present spontaneous mutations and recombinations of the Apaf-1 gene and its<br />neighbouring sequences. We sequence 48 colonies containing pcDNA3.1 vector with<br />Nluc/Apaf1 and Cluc/Apaf1 obtained through the quick-change site-directed mutagenesis<br />method, transforming to DH5-α and XL10-Gold at two temperatures, 18 and 37ºC. In 21 of<br />these cases, we found 38 different mutations. Our data suggest that there is a direct relationship<br />between bacterial incubation temperatures and the number of unwanted spontaneous mutations.<br />During our experiment we found that the Apaf-1 gene is much less susceptible to spontaneous<br />mutations when it is transformed into XL10-Gold at 18 ºC . In contrast, a large number of<br />spontaneous mutations were found when the gene of interest transformed into DH5α at 37ºC .<br /><br /> https://pbiosci.ut.ac.ir/article_52304_ce8f33ace899b4c12db1452d67f5fe75.pdf