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EFFICIENT METHOD OF PREPARING DNA INVERTED REPEAT STRUCTURE

机译:DNA反向重复结构的高效制备方法

摘要

Conventional techniques for preparing a chimera gene having an inverted repeat sequence of a target sequence suffer from complications since a target sequence is inserted into 2 sites on a vector in sense and antisense orientations, and restriction enzyme recognition sequences must be independently provided at an insertion site of a vector and both ends of the target sequence.;In this invention, a cassette construct comprising an arbitrary adaptor sequence and an inverted adaptor sequence separated by an arbitrary spacer sequence or a plasmid vector comprising such cassette construct incorporated therein is prepared, a target sequence is ligated to either or both ends of the cassette construct, or a target sequence is inserted into one end of the cassette construct on the plasmid vector, followed by PCR. Thus, a chimera gene having an inverted repeat sequence is prepared.
机译:用于制备具有靶序列的反向重复序列的嵌合基因的常规技术存在复杂性,因为靶序列以有义和反义方向插入到载体的2个位点中,并且必须在插入位点处独立提供限制性内切酶识别序列。在本发明中,制备包含被任意间隔子序列隔开的任意衔接子序列和反向衔接子序列的盒构建体或包含掺入其中的这种盒构建体的质粒载体,制备靶将序列连接至盒构建体的任一端或两端,或将靶序列插入质粒载体上的盒构建体的一端,然后进行PCR。因此,制备了具有反向重复序列的嵌合基因。

著录项

  • 公开/公告号EP1672064A4

    专利类型

  • 公开/公告日2006-12-20

    原文格式PDF

  • 申请/专利权人 RIKEN;

    申请/专利号EP20040773482

  • 发明设计人 FUKUDA HIROO;DEMURA TAKU;

    申请日2004-09-22

  • 分类号C12N15/09;C12N1/19;C12N1/21;C12N5/10;C12N15/10;C12N15/11;C12N15/66;C12N15/70;

  • 国家 EP

  • 入库时间 2022-08-21 20:51:04

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