摘要
可转化人工染色体 (Transformation competentArtificialChromosome ,TAC)是具有克隆和转移大片段基因能力的新型载体 ,是植物基因克隆和转化的有效工具。为了克隆小麦抗白粉病基因和其它基因 ,本研究用TAC载体 pYLTAC17构建了带有抗白粉病基因Pm2 1的小麦 簇毛麦 6VS/6AL易位系的基因组DNA文库。该文库包含2 10万个克隆 ,平均插入片段 35kb ,库容相当于普通小麦基因组的 4 9倍。本文库以约 10 0 0个克隆组成 1个混合池的方式保存在 2 2块 96孔平板 ,可用PCR方法进行文库的筛选。
Transformation\|competent artificial chromosome (TAC) vector is able to clone and transfer large DNA fragments in plants and is a powerful tool for plant gene isolation and transformation.To clone important genes from wheat,a TAC genomic library was constructed from nuclear DNA of a 6VS/6AL wheat\| Haynaldia villosa translocation line that harbor the gene Pm21 for resistance to powdery mildew.The library consists of 2 1×10 6 clones with an average DNA insert size of 35kb,and represents in total 4.9 genome equivalents.The library was stored as clone pools in 96\|well plates,and each pool contained about 1000 clones.TAC clones containing gene(s) of interest can be screened by a pooled\|PCR/colony\|hybridization strategy.
出处
《生物工程学报》
CAS
CSCD
北大核心
2000年第4期433-436,共4页
Chinese Journal of Biotechnology
基金
国家高技术研究发展计划项目资助!(Z 17 0 4 0 1)
关键词
可转化人工染色体
基因组文库
小麦
基因载体
Transformation\|competent artificial chromosome,genomic DNA library,wheat
