文档介绍：
野生稻基因组 PCR 扩增体系的探索和优化#
欧阳亦聃*
(华中农业大学生命科学技术学院,武汉,430070)
5
10
15
20
25
30
35
40
摘要:本研究运用 CTAB 法提取野生稻的基因组 DNA,用作 PCR 模版,根据粳稻日本晴基
因组序列中 S5 区段设计多对引物,进行 PCR 扩增和条件优化。结果发现覆瓦式 PCR 引物
以及多轮 PCR 反应能获得清晰明亮的 PCR 扩增带型。基于基因编码区序列所设计的引物扩
增效率较高。扩增产物的 GC 含量较高时,在 PCR 体系中加入 2%DMSO 能有效提高扩增效
率。
关键词:分子生物学;野生稻;PCR 反应条件优化
中图分类号:Q756
Investigation and optimization of the PCR amplification
system for the wild rice genome
OUYANG Yidan
(The College of Life Science and Technology, Huazhong Agricultural University, Wuhan,
430070)
Abstract: The wild rice genomic DNA was extracted by CTAB method and was further used as
the PCR template. The PCR primers were designed based on the S5 region of japonica
Nipponbare genome sequence. Experiments were conducted to optimize PCR amplification. The
imbricate PCR primers and multiple rounds of reactions can increase the PCR amplification
efficiency and obtain clear and bright bands. The primers based on the coding region sequence of
the protein-encoding gene were more preferably than other primers. For the genomic sequence
with higher GC content, adding 2% DMSO in PCR system will improve the amplification
efficiency.
Key words: Molecular biology; wild rice; optimization of PCR
0 引言
水稻是我国主要粮食作物,对国民经济、粮食安全具有举足轻重的意义。稻属植物含十
个基因组类型的水稻种,包括亚洲栽培稻和非洲栽培稻两个栽培稻种,和 20 多个野生稻种
[1]
籼稻和粳稻之间存在生殖隔离现象即两个亚种的杂交后代育性较低,这种杂种不育现象是植
物生殖隔离的代表性实例[2~4]。最近通过图位克隆