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۱Mutating Asn–666 to Glu in the O–helix region of the taq DNA polymerase gene
اطلاعات انتشار: Research in Pharmaceutical Sciences، پنجم،شماره۱(پياپي ۹)، ۲۰۱۰، سال
تعداد صفحات: ۵
Taq DNA polymerase is widely used in laboratories and for this reason many investigators have focused their attention on understanding the role of various regions and amino acids in this enzyme. O–helix is a part of taq polymerase suggested to play an important role in the enzyme fidelity. The influence of Asn666 in this helix on the enzyme function has never been investigated, and therefore by using nested PCR, a portion of taq DNA polymerase gene containing Asn666Glu mutation was amplified. This DNA was digested with Eco RI restriction enzyme to confirm the presence of Asn666Glu mutation. After digesting this product and the wild type taq–pET–15b plasmid with NheI and BamHI restriction enzymes, they were ligated and used for the transformation of E. coli DH5α competent cells. The obtained colonies were screened for the presence of the mutated taq polymerase gene using EcoRI, NdeI and BamHI restriction enzymes. In conclusion, with the use of the obtained recombinant plasmid it is possible to study the role of this amino acid on taq DNA polymerase function.

۲Genotoxic effects of some l–[(benzofuran–2–yl)–phenylmethyl]–imidazoles on MCF–7 cell line
اطلاعات انتشار: Research in Pharmaceutical Sciences، هفتم،شماره۳(پياپي ۱۵)، ۲۰۱۲، سال
تعداد صفحات: ۸
Increased exposure to estrogen has been associated with the risk of breast cancer. Substituted benzofuran derivatives with inhibitory effects on estrogen synthesis could be considered as a potential approach to reduce the risk of breast cancer. The study of cytotoxic effects of these compounds has suggested involvement of other mechanisms such as DNA damage. In the current study we have investigated genotoxic effects of some benzofuran derivatives on MCF–7 cell line. The MCF–7 cell line was exposed both to benzofuran phenylmethyl imidazole and its 4– fluoro, 4–chloro, 2–methoxy and 2–methyl derivatives for 2 h. The Comet assay was used to examine DNA damage due to this exposure. We also studied the DNA repair capacity after 2 h exposure to genotoxic concentrations of these compounds and their recovery were evaluated after 17 and 24 h, using the comet assay. The results indicated that genotoxic effects of these compounds appeared in concentrations of 10–8 to 10–6 M. The 4– fluoro and 4–chloro derivatives exhibited the highest genotoxicity and the unsubstituted benzofuran phenylmethyl imidazole had the lowest effect. The 4– fluoro, 4–chloro and 2–methyl derivatives were recovered after 24 h while 2–methoxy and the unsubstituted derivatives were recovered after 17 h. The results showed that these compounds are genotoxic and the concentration of tested benzofuran derivatives with genotoxic effects are not close to their enzyme inhibitory concentration. Moreover, our study shows that the DNA damages are repairable. Therefore, it seems that the investigated compounds have the potentials as therapeutic agents.
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