摘要
从未成熟的大豆子叶中提取总RNA，利用RT－PCR方法扩增出大豆Kunitz型胰蛋白酶抑制剂基因单一目的片段，克隆到pBluescriptKS（＋）SmaI位点上。序列分析表明，该基因与报导的序列高度同源：其核苷酸序列及推论的编码氨基酸序列的同源率分别为99．5％和98．2％。由此，构建了大豆Kunitz型胰蛋白酶抑制剂基因的植物高效表达载体pBinSKTI。并采用农杆菌介导的叶盘法转化了烟草，获得的转基因烟草再生植株经抗虫测试表明：与对照烟草相比，具有明显的抗棉铃虫能力。
The Soybean Kunitz Trypsin Inhibitor gene was obtained from soybean immature cotyledons total RNA by RT-PCR, and the amplified DNA fragment was cloned into Smal site of pBluescript KS (+). DNA sequence analysis indicates that the cloned DNA fragment composed of 663hp has 99. 5% homologies in nucleotide sequence and 98. 2% in amino acid sequence respectively compared with previously published sequence. A plant expression vector pBinSKTI was constrncted and used in plant transformation. The transgenic tobacco was confirmed by PCR and scutnern blotting. Results of Bioassays show that the transgenic tobacco displays notable insectrestnnt ability to the Larvae of Heliothis armigera compared with wild tobacco.
出处
《高技术通讯》
EI
CAS
CSCD
1997年第9期5-9,共5页
Chinese High Technology Letters
基金
863计划
"总理基金"
美国"洛氏基金"
关键词
大豆胰蛋白酶
抑制剂
抗虫基因
转基因烟草
Soybean Kunitz Trypsin Inhibitor, RT-PCR, Analytical sequencing, Insecticidal gene, Transgenic tobacco
