专利快速检索-快速检索全球专利，免费商用专利数据库-IPRDB

1. 发明授权

US5994528A Method of detecting gene expression and/or of preventing such expression in cells 失效
标题翻译：在细胞中检测基因表达和/或预防这种表达的方法
公开(公告)号：US5994528A
公开(公告)日：1999-11-30
申请号：US38014
申请日：1998-03-11
申请人： Thuraiayah Vinayagamoorthy , Eric Schloss
发明人： Thuraiayah Vinayagamoorthy , Eric Schloss
IPC分类号： C12Q1/68 , C07H21/02 , C07H21/04 , C12P19/301
CPC分类号： C12Q1/6851
摘要： A method of amplifying a nucleotide sequence complementary to an mRNA template derived from genomic DNA. The method involves the following steps. A sample mixture containing an mRNA template and corresponding genomic DNA is provided, the genomic DNA including at least two exons separated by at least one intron. A pair of intron-blockers are introduced into the mixture, the intron-blockers comprising a sequence of intron-specific oligonucleotides modified to prevent nucleotide extension in conditions promoting polymerase chain reaction. A primer pair promoting amplification of cDNA derived from the mRNA template is introduced into the mixture and then reverse transcription polymerase chain reaction is carried out to amplify cDNA. Detection of the cDNA is proof of the existence of mRNA in the sample, and thus proof of expression of the corresponding gene. The method avoids false positives caused by amplification of genomic DNA as well as cDNA based on an mRNA template. The invention includes a method of suppressing gene expression in vivo, which comprises exposing cells containing a gene to be suppressed, made up of exons and at least one intron, to intron-blockers having nucleotide sequences that bind to the intron to prevent gene expression.
摘要翻译：扩增与衍生自基因组DNA的mRNA模板互补的核苷酸序列的方法。该方法包括以下步骤。提供了含有mRNA模板和相应的基因组DNA的样品混合物，所述基因组DNA包括至少两个由至少一个内含子分开的外显子。将一对内含子阻断剂引入混合物中，内含子阻断剂包含修饰以在促进聚合酶链反应的条件下防止核苷酸延伸的内含子特异性寡核苷酸序列。将从mRNA模板衍生的cDNA扩增的引物对引入混合物中，然后进行逆转录聚合酶链反应以扩增cDNA。 cDNA的检测证明了样品中mRNA的存在，从而证明了相应基因的表达。该方法避免了基因组DNA扩增引起的假阳性以及基于mRNA模板的cDNA。本发明包括抑制体内基因表达的方法，该方法包括将含有由外显子和至少一个内含子组成的待抑制基因的细胞暴露于具有与内含子结合的核苷酸序列以防止基因表达的内含子阻断剂。

IPRDB

热门服务

关于我们

友情链接

联系方式