摘要
以2,4-D处理的刺五加体细胞胚为实验材料,提取总RNA并分离mRNA合成cDNA,连接到pAP3neoPredigested载体上。采用电穿孔法将重组质粒转化到DH10B中。经测定,原始文库滴度为2.3×106pfu·mL-1,重组率大于95%。插入片段的长度大部分在0.5～2.0kb之间,平均长度为0.96kb,表明刺五加体细胞胚cDNA文库已构建成功。
Total RNAs were extracted from 2,4-D pretreated Eleutherococcus senticosus somatic embryos, cDNAs were synthesized from isolated mRNAs and ligated into the pAP3neo Predigested vector. The recombinant plasmids were transformed into Escherichia coli DH10B by electroporation. The library qualification evaluation results showed that the titer was larger than 2.3× 10^6 pfu·mL^-1, the percentage of recombination 〉95%, the size of most inserted fragments ranged between 0.5 kb and 2.0 kb, the average size was 0.96 kb. It indicated that E. senticosus somatic embryo cDNA library was constructed successfully.
出处
《植物生理学通讯》
CAS
CSCD
北大核心
2008年第4期715-718,共4页
Plant Physiology Communications
基金
国家自然科学基金(30671701)
关键词
刺五加
体细胞胚发生
CDNA文库
Eleutherococcus senticosus
somatic embryogenesis
cDNA library
