摘要
为了寻找一种简捷的克隆同源基因的途径,以小麦铁蛋白基因的同源克隆为例,利用玉米的铁结合蛋白基因序列比对小麦的EST库,在网上进行电子延伸后以起始密码子为起点,终止密码子为终点设计特异性强的引物进行基因克隆,经过引物O1和Fcds扩增片段的测序及Blast比对,证实本试验已成功地从小麦的基因组和cDNA中克隆出铁结合蛋白基因。最后对基于同源序列克隆基因的策略及引物设计技巧进行了探讨。
The development of comparative genomics makes homologous cloning an important method to discover new gene. To find an efficient method that fit for homologous gene cloning, we studied the strategy of gene cloning and skill of primers design based on homologous sequence, taking the ferritin gene as an example. The EST databank of wheat was hlased with the sequence of ferritin gene from maize at the first,then the gene was cloned in silico. Based on the result of in silico cloning, the ferritin genes were cloned from wheat with the special primers which designed from the start code region and the terminator region. Ferritin genes were cloned successfully from genomic DNA and cDAN of wheat. The method to clone genes based on homologous sequence from wheat was discussed on the strategy and skill of primers design.
出处
《麦类作物学报》
CAS
CSCD
北大核心
2008年第1期25-30,共6页
Journal of Triticeae Crops
基金
国家“973”项目(2004CB117200)
河南工业大学博士基金项目(150216)
关键词
小麦
铁结合蛋白基因
引物设计
同源克隆
Gene cloning based on homologous sequence
Primer design
Ferritin
