青蒿素的瞬时转化

Dongming Ma; Hong Wang

doi:10.21769/BioProtoc.1476

Improve Research Reproducibility A Bio-protocol resource

提交稿件
订阅
CN
- EN - English
- CN - 中文

Peer-reviewed

Transient Transformation of Artemisia annua

青蒿素的瞬时转化

Dongming Ma

Hong Wang

发布: 2015年05月20日第5卷第10期 DOI: 10.21769/BioProtoc.1476 浏览次数: 8340

评审: Tie LiuAnonymous reviewer(s)

下载 PDF

Q&A

引用

Cited by

参见作者原研究论文

The authors used this protocol in:

Cover of Plant & Cell Physiology, featuring study using the protocol.

Sep 2014

实验方案合集

Cell Imaging - A Special Collection for Cell Bio 2023

相关实验方案

双向双流式根芯片在不对称胁迫处理下对植物主根的生理分析

Claudia Allan [...] Claudia-Nicole Meisrimler

2023年08月05日 1077 阅读

利用分裂可选择标记的植物转化新型基因叠加方法

Guoliang Yuan [...] Xiaohan Yang

2025年02月20日 962 阅读

甜樱桃发育中花原基的淀粉酶法定量检测

Nestor Santolaria [...] Afif Hedhly

2025年04月05日 805 阅读

Abstract

Transient transformation of Artemisia annua does not depend on chromosomal integration of heterologous DNA, and recombinant DNA can be introduced into plant cells via Agrobacterium aided by vacuum. The leaves of 7th and 8th internode from 4-week-old seedlings were chosen as explants, a vacuum system was applied to facilitate agrobacteria into plant cells, the co-cultivation was in the dark at 25 °C for 36-72 h, then GUS or GFP maker genes were used for testing the efficiency of the transformation. The method is used for quick transferring of genes into Artemisia annua (A. annua) by transient transformation.

Keywords: Artemisia annua (阿尔泰米夏黄花)

Agrobacterium tumefaciens (农杆菌介导法)

Transient transformation (瞬时转化)

Materials and Reagents

Artemisia annua leaves from 4-week-old aseptic seedlings
Agrobacterium tumefaciens (A. tumefaciens) strain EHA105 (or LBA4404)
Lysogeny broth (LB) medium (pH 7.0, containing 50 mg/L rifampicin and 50 mg/L kanamycin)
Murashige and Skoog medium (MS) liquid medium (Sigma-Aldrich, catalog number: M5524 )
100 μM acetosyringone (Sigma-Aldrich, catalog number: D134406 )
10 mM MgCl₂
0.005% Silwet L-77 (Agri-Turf Supplies, catalog number: VIS-01 )
Glycerol
Filter paper (Lab Depot, catalog number: TLDCFP1-032 )
Yeast extract broth (YEB) medium (see Recipes)