摘要
从大豆 (花生豆 1号 )未成熟种子中提取总RNA ,逆转录形成cDNA第一条链。根据genebank中大豆 11S球蛋白Gy5的cDNA序列设计引物 ,用PCR方法扩增Gy5的cDNA序列 ,克隆到 pUC19质粒载体上 ,并测定其全序列。用限制性内切酶PstI和EcoRI酶切重组质粒 ,将目的片段与质粒 pCAMBIA130 1连接 ,构建成植物表述载体并转化到农杆菌中 ,以便于以后的研究利用。
The total RNA was estracted from immature seeds of soybean and directly used in single-stranded(ss)DNA synthetic reaction. According to soybean 11S glycinin Gy5 cDNA sequences in genebank,a pair of primers were designed.Double-stranded(ds)cDNA was amplified by PCR?The amplified Gy5 cDNA was cloned into plasmid pUC 19 and sequenced?Fragment of the Gy5 cDNA in the recombinant plasmid was cut with PstI/EcoRI and ligated with plasmid Pcambia1301,then a plant expression vector was constructed?It was also transferred into Agrobacterium LBA4404 in order to be utilized later.
出处
《大豆科学》
CAS
CSCD
北大核心
2004年第3期159-163,共5页
Soybean Science
基金
"国家转基因植物中试与产业化基地 (吉林 )"专项课题 (J99-B -0 0 1)
