《植物生理学报》 2012, 48(1): 95-101

小金海棠MxYSL5启动子克隆技术的比较

陈竹, 王忆, 殷丽丽, 张新忠, 韩振海*

中国农业大学园艺植物研究所, 北京100193

通信作者：韩振海；E-mail: rschan@cau.edu.cn；Tel: 010-62732467

摘　要：

分别利用3种基于PCR技术的染色体步移法克隆了小金海棠MxYLS5基因的启动子, 并比较了这三个体系的扩增产物。3种体系都可以获得特异性扩增产物, 但扩增条带的长度和数量均不同, 其中利用Genome Walking Kit扩增出来的条带特异性较好长度最长, 而且特异性引物和简并引物的设计及退火温度的设置对3种体系的扩增产物起着关键的作用。基于PCR技术的染色体步移法为小金海棠MxYLS5基因启动子的克隆提供了一个稳定可靠的技术手段。

关键词：启动子克隆; 染色体步移; Tail-PCR; hiTail-PCR; 小金海棠

收稿：2011-09-26   修定：2011-12-05

资助：现代农业产业技术体系专项资金(CARS-28)、北京市科技新星课题(2008B74)和果树逆境生理与分子生物学北京市重点实验室资金。

Comparision of MxYSL5 Promoter Cloning Technology from Malus xiaojinensis

CHEN Zhu, WANG Yi, YIN Li-Li, ZHANG Xin-Zhong, HAN Zhen-Hai*

Institute for Horticultural Plants, China Agricultural University, Beijing 100193, China

Corresponding author: HAN Zhen-Hai; E-mail: rschan@cau.edu.cn; Tel: 010-62732467

Abstract:

Three genomic walking methods based on PCR were used to clone the promoter of MxYLS5 from Malus xiaojinensis. And we compared the amplification products of these three systems. All these systems can obtain specific amplification products, but the length and numbers of bands are different. Amplified bands applied Genome Walking Kit have better specificity and longer length. And specific primers, degenerate primers and annealing temperature play a key role in amplification products. These three systems provide a stable and reliable technological means for promoter cloning of MxYLS5 from Malus xiaojinensis.

Key words: promoter cloning; genomic walking; Tail-PCR; hiTail-PCR; Malus xiaojinensis