摘要
以野生罂粟(Papaver somniferum)试管苗幼叶为材料,用Trizol试剂盆提取总RNA,通过RT-PCR法获得BBE基因的cDNA片段,序列分析表明,所获得的cDNA序列全长1 608 bp,具有完整的开放阅读框(ORF),编码536个氨基酸,经blast检索该片段与GenBank中的小檗碱桥酶基因BBE(AF025430)同源性为94.84%.以中间载体pHANNIBAL和植物表达载体pART27为基础,构建了CaMV-35S启动子驱动的含小檗碱桥酶基因片段反向重复序列的RNAi双元表达载体pARB,为培育低吗啡,高蒂巴因的罂粟新品系奠定了基础.
Total RNA was extracted from young leaf of opium poppy （Papaver somniferum L） by using Trizol regent. The berberine bridge enzyme（BBE） gene was cloned by RT- PCR and its coding sequence was analyzed. The result indicated that the full-length cDNA was 1 608 bp and contained a 1 608 bp open reading frame （ORF） encoding 536 amino acids. The sequence analyzed with software Blast was highly homologous to the BBE（AF025430）gene with a 94. 84 % identity to it. Then the promoter CaMV 35S driven, plant siRNA expression vector pARB with inverted repeats of BBE was constructed based on the vectors pHANNIBAL and pART27. The work will lay the foundation for breeding a low morphine and high thebaine poppy.
出处
《甘肃农业大学学报》
CAS
CSCD
北大核心
2009年第6期52-56,106,共6页
Journal of Gansu Agricultural University
基金
教育部博士学科点研究基金(2007733008)
甘肃省卫生厅重点中医药科技项目(2008HDTC02)
甘肃省卫生厅重点中医药科技项目(GZK-2009-10)
关键词
罂粟
BBE基因
RNAI载体构建
克隆
Papaver somniferum
berberine bridge enzyme gene
construction of RNAi plant expression vector
clone
