利索-全球专利检索

  1. 登录
  2. 注册
发明公开
CN104195142A 一种香石竹PS1基因ihpRNA载体的构建方法 无效 - 驳回

一种香石竹PS1基因ihpRNA载体的构建方法
摘要：
本发明公开了一种香石竹PS1基因ihpRNA表达载体的构建方法，本发明首次从香石竹中获得一个与2n配子形成相关的PS1基因，该基因cDNA全长3423bp，编码1140个氨基酸。以香石竹DNA为模板，扩增PS1基因466bp外显子和85bp内含子序列，然后扩增466bp外显子反义片断，将正义和内含子片断以及反义片断同时插入到pCAMBIA1301-220.6表达载体中，从而构成反向互补发卡结构的RNAi(ihpRNA)表达载体，本发明简化了构建的步骤，且无需专门的RNAi载体，具有简单、快速、高效和成本低等优点，为大规模的植物功能基因研究提供了一个高通量的平台。
摘要（英）：

The invention discloses a method for building an ihpRNA carrier of a Dianthus caryophyllus PS1 gene. According to the method, a PS1 gene related to 2n gamete formation from Dianthus caryophyllus is obtained for the first time, wherein the cDNA overall length of the gene is 3423bp, and the gene is used for encoding 1140 amino acids. The method comprises the following steps: by taking Dianthus caryophyllus DNA as a template, amplifying 466bp exon and 85bp intron sequences of the PS1 gene, and then amplifying a 466bp exon antisense fragment; and simultaneously inserting sense and intron fragments and an antisense fragment into a pCAMBIA1301-220.6 expression carrier, so as to form an RNAi (ihpRNA) expression carrier of a reverse complementary hairpin structure. According to the method, the building steps are simplified, a special RNAi carrier is not needed, the method has the advantages of being simple, fast, high in efficiency, low in cost and the like, and a high throughput platform is provided for large-scale research of plant functional genes.

0/0