公开(公告)号：US20110130304A1

公开(公告)日：2011-06-02

申请号：US12675953

申请日：2008-08-29

申请人： Kai Rauasalu ,  Anna Iofik ,  Valeria Lulla ,  Liis Karo-Astover ,  Kristi Tamm ,  Liane Ulper ,  Inga Sarand ,  Andres Merits

发明人： Kai Rauasalu ,  Anna Iofik ,  Valeria Lulla ,  Liis Karo-Astover ,  Kristi Tamm ,  Liane Ulper ,  Inga Sarand ,  Andres Merits

IPC分类号： C40B40/02 ,  C40B50/06 ,  C40B40/06 ,  C07H21/04 ,  C12N15/63

CPC分类号： C12N15/86 ,  C12N15/1027 ,  C12N15/1034 ,  C12N2770/36143

摘要： A method for creating an alphavirus-based genomic library, comprising a) ligation of foreign sequence (s) from an expression library or a random library into plasmids containing cloned alphaviral cDNA, b) multiplication of the obtained plasmid constructs in bacterial cells, c) direct transfection of the obtained plasmid constructs into mammalian or arthropod cells, characterized in that the sequence of an intron or sequences of introns are inserted into the respective genome of an alphavirus or into the cDNA of an expression vector based on an alphavirus, —the sequence of a viral subgenomic promoter, which is larger than minimal functional promoter is inserted immediately to the 3′ end of the sequences coding the structural proteins of the named alphavirus, —and ribozyme sequence is inserted for creating correct 3′ ends of the alphavirus.

摘要翻译： 一种用于产生基于甲病毒病毒的基因组文库的方法，包括：a）将外源序列从表达文库或随机文库连接到含有克隆的甲病毒cDNA的质粒中，b）将获得的质粒构建体在细菌细胞中增殖，c） 将获得的质粒构建体直接转染到哺乳动物或节肢动物细胞中，其特征在于将内含子或内含子序列的序列插入到甲病毒病毒的相应基因组或基于甲病毒属的表达载体的cDNA中，该序列 将大于最小功能性启动子的病毒亚基因组启动子立即插入到编码所述甲病毒属的结构蛋白的序列的3'末端，并插入核酶序列以产生甲病毒的正确3'末端。