摘要
从甘农薯1号马铃薯试管苗叶片中提取总DNA,根据Genebank报道的马铃薯多酚氧化酶基因 POT32 序列,设计合成了带有BamH 、Sac 特定酶切位点的2对特异引物,通过PCR获得1840bP和476bP的扩增产物.测序结果表明,1840bp的扩增产物与Genebank中发表的序列一致,476bp的扩增产物正是欲扩增的马铃薯多酚氧化酶基因的同源片段.将2个扩增产物反向连接到表达载体PC3中,通过双酶切鉴定后,按直接导入法实现反义基因表达载体质粒向农杆菌EHA105的导入;并经PCR鉴定证明,此质粒已整合到农杆菌Ti上.
In present studies,the general DNA was extracted from the potato leaves of 'Gannong No.1'.Two pairs of special primer were designed according to POT32 sequence by Genebank published,and two amplification products were obtained (one is 1 840 bp,another is 476 bp).Antidirect linked them to the PC3 expression vector,and checked them with two enzymes cutting.Then made the antisense expression vector into Agrobacterium EHA105 by the method of directing translation.The conclusion was that vector have been into EHA105.
出处
《西北植物学报》
CAS
CSCD
2003年第10期1745-1749,共5页
Acta Botanica Boreali-Occidentalia Sinica
基金
国家高技术研究发展计划 " 863"计划项目(2001AA241131 )
关键词
马铃薯
PPO基因克隆
反义基因表达载体构建
potatoes
polyphenol oxidase gene clone
anti-sense expression vector construction
