摘要
根据GenBank中大白菜(Brassica campestris L. ssp. pekinensis)基因组测序结果,分析了大白菜PAT1(phytochrome A signal transduction)基因的内含子和外显子。通过拼接外显子,获得了大白菜PAT1基因的cDNA编码序列。根据编码区序列,设计并合成了一对引物;利用这对引物,采用RT-PCR技术扩增出大白菜福山包头PAT1基因(CcPAT1)的全长cDNA序列。CcPAT1编码区共1 500 bp,与拟南芥PAT1相似性为85.03%。同时构建了含有35S启动子的pCAMBIA-2300-CcPAT1过表达载体,并利用PAT1 5′端特异区域构建了pFGC-1008-PAT1RNAi(RNA interference)载体,以备将来研究大白菜CcPAT1基因的功能。
The intron and exon of PAT1 （phytochrome A signal transduction） gene in Chinese cabbage （ Brassica campestris L. ssp. pekinensis ） were. analyzed based on the genomic DNA sequence in Gen- Bank. By splicing the exons, the cDNA sequence of PAT1 gene was obtained. Then a pair of primers was designed for RT - PC1L The results showed that the full - length CcPAT1 cDNA of Chinese cabbage variety Fushanbaotou was isolated, which was 1 500 bp and 85.03% homologous to the Arabidopsis thaliana PAT1 gene. To study the functions of CcPAT1 , the overexpression vector of pCAMBIA -2300 - CcPAT1 with 35S promoter, and the PAT1 RNA interference expression vector of pFGC - 1008 - PATI with the special 5＇ - terminal sequence of PAT1 were constructed.
出处
《山东农业科学》
2009年第2期1-4,共4页
Shandong Agricultural Sciences
基金
国家科技基本条件平台建设项目(2005DKA21001-38)
山东省农业科学院高技术自主创新基金项目(2007YCX003)
