期刊论文

Construction of CRISPR/Cas9 knockout vector BnaSDG8 and its genetic transformation in Brassica napus

中文

湖南农业大学学报(自然科学版)

1007-1032

2018

44

4

346-352

10.13331/j.cnki.jhau.2018.04.002

作物种质创新与资源利用国家重点实验室科学基金开放项目

本校为第一机构

以甘蓝型油菜‘湘油15号’为试材,通过生物信息学分析,运用CRISPR/Cas9系统对Bna SDG8第1个外显子上的Bna SDG8–A和Bna SDG8–C保守序列区域进行CRISPR/Cas9敲除位点设计,构建了CRISPR/Cas9敲除载体p CAMBIA1300–sg RNA/Cas9–Bna SDG8;通过遗传转化,获得转化植株19株;经测序比对,4株植株的Bna SDG8被成功编辑;通过对定向敲除植株开花时间的统计分析,初步确定Bna SDG8参与甘蓝型油菜开花时间的调控。

In this study, Brassica napus L. ‘Xianyou 15＇ was used as the test material. By bioinformatics analysis, the CRISPR/Cas9 knockout sites were designed for the Bna SDG8–A and Bna SDG8–C conserved sequence regions on the first exon of Bna SDG8. The CRISPR/Cas9 knockout vector p CAMBIA1300–sg RNA/Cas9–Bna SDG8 was successfully constructed; 19 transformed plants were obtained through genetic transformation, 4 plants were successfully edited in 19 transformed plants. In addition, through the statistical analysis of flowering time of directional...