TY - JOUR ID - 52304 TI - Apoptotic protease-activating factor 1 (Apaf-1) as a liable gene for spontaneous mutations in vitro JO - Progress in Biological Sciences JA - PBS LA - en SN - 1016-1058 AU - Azad, Taha AU - Tashakor, Amin AU - Ghahremani, Mina AU - Hemmati, Roohullah AU - Ataei, Farangis AU - Hosseinkhani, Saman AD - Department of Biochemistry, Faculty of Biological Sciences, Tarbiat Modares University, Tehran, Iran AD - Department of Plant Biology, Faculty of Biological Sciences, Tarbiat Modares University, Tehran, Iran Y1 - 2014 PY - 2014 VL - 4 IS - 2 SP - 261 EP - 273 KW - Apoptosis KW - apoptotic protease-activating factor 1 (Apaf-1) KW - cancer KW - spontaneous mutations KW - WD-40 repeats DO - 10.22059/pbs.2014.52304 N2 - The apoptotic protease-activating factor 1 (Apaf-1) receives the death signal in the intrinsic ormitochondrial pathway of apoptosis. Upon the releasing of cytochrome c from theintermembrane space of mitochondria and binding to Apaf-1 molecules, a heptamericapoptosome complex is formed and triggers the downstream cascade of caspases. Here, for thefirst time we present spontaneous mutations and recombinations of the Apaf-1 gene and itsneighbouring sequences. We sequence 48 colonies containing pcDNA3.1 vector withNluc/Apaf1 and Cluc/Apaf1 obtained through the quick-change site-directed mutagenesismethod, transforming to DH5-α and XL10-Gold at two temperatures, 18 and 37ºC. In 21 ofthese cases, we found 38 different mutations. Our data suggest that there is a direct relationshipbetween bacterial incubation temperatures and the number of unwanted spontaneous mutations.During our experiment we found that the Apaf-1 gene is much less susceptible to spontaneousmutations when it is transformed into XL10-Gold at 18 ºC . In contrast, a large number ofspontaneous mutations were found when the gene of interest transformed into DH5α at 37ºC . UR - https://pbiosci.ut.ac.ir/article_52304.html L1 - https://pbiosci.ut.ac.ir/article_52304_ce8f33ace899b4c12db1452d67f5fe75.pdf ER -