Olafsdóttir Gudbjörg, Svansson Vilhjálmur, Ingvarsson Sigurdur, Marti Eliane, Torsteinsdóttir Sigurbjörg
Institute for Experimental Pathology, University of Iceland, Keldur, Reykjavík, Iceland.
Acta Vet Scand. 2008 Nov 4;50(1):44. doi: 10.1186/1751-0147-50-44.
One of the prerequisite for developing DNA vaccines for horses are vectors that are efficiently expressed in horse cells. We have analysed the ectopic expression of the human serum albumin gene in primary horse cells from different tissues. The vectors used are of pcDNA and pUC origin and include the cytomegalovirus (CMV) promoter. The pUC vectors contain CMV intron A whereas the pcDNA vectors do not. Insertion of intron A diminished the expression from the pcDNA vectors whereas insertion of a Kozak sequence upstream of the gene in two types of pUC vectors increased significantly the in vitro expression in primary horse cells derived from skin, lung, duodenum and kidney. We report for the first time the significance of full consensus Kozak sequences for protein expression in horse cells in vitro.
开发马用DNA疫苗的前提条件之一是能在马细胞中高效表达的载体。我们分析了人血清白蛋白基因在来自不同组织的原代马细胞中的异位表达。所使用的载体源自pcDNA和pUC,包括巨细胞病毒(CMV)启动子。pUC载体含有CMV内含子A,而pcDNA载体则没有。内含子A的插入降低了pcDNA载体的表达,而在两种pUC载体的基因上游插入Kozak序列则显著提高了源自皮肤、肺、十二指肠和肾脏的原代马细胞中的体外表达。我们首次报道了完全一致的Kozak序列对马细胞体外蛋白质表达的重要性。