کلونینگ و بیان موقت ژن محافظت‌کننده سلولی، HO-1 ، در سلول‌های بنیادی مزانشیمال از طریق سیستم بیانی آدنوویروس بر مبنای تکنولوژی Gateway

Abstractbackground and objectivesheme oxegenase1 (ho-1) is one of the potent cytoprotective factors. the goal of this study was to perform cloning and transient over expression of the human ho-1 gene in mesenchymal stem cells (mscs) using the adenoviral expression system based on the gateway technology.materials and methodsin order to induce expression of ho-1, a549 cell lines were exposed to uv for 1 hour. the full length cdna of ho-1 was isolated and cloned into pentr topo/d vector by topo cloning reaction. to construct the expression clone, a lr recombination reaction was carried out between the entry clone and destination vector, pad/cmv/v5-dest. the recombinant virus was produced in the appropriate mammalian cell line. mscs were infected by the recombinant virus expressing ho-1.resultsthe results showed that human recombinant ho-1 was successfully cloned and the accuracy of the gene and its frame in the vector were confirmed by dna sequencing. expression of ho-1 in mscs was confirmed by rt-pcr and western blot analysis. the results indicated that the expression of ho-1 is transient.conclusions transient expression of human ho-1 gene in mscs by using adenovirus expression system may be considered as an efficient gene transfer strategy into mscs in order to promote stem cell therapy.