甘蓝型油菜中黑芥子酶基因在大肠杆菌中重组质粒的构建和表达

实验通过RT-PCR程序从提取的油菜总RNA中扩增出硫代葡萄糖苷水解酶（又称黑芥子酶，EC3.2.1.147）基因,酶解后插入大肠杆菌表达载体（Escherichia coli）pGEX-4T-1，获得克隆菌株。基因测序在 GenBank中的登陆号为EF583560，翻译的氨基酸序列与已报道的黑芥子酶（GenBank中的登陆号为Q00326）中的一段有一个氨基酸不同，同源性达到99%。经过IPTG诱导表达，在91KDa左右处有表达量很高的一条带。

Construction and Expression of a Brassica napus Myrosinase in Escherichia coli

The myrosinase gene was amplified from total RNA by RT-PCR and inserted into Escherichia coli expression vector pGEX-4T-1.The result of sequencing was EF583560 in GenBank.The amino acid sequence showed that the homology was 99% comparing with a length of published amino acid sequence of Q00326.The recombinant protein was highly expressed at 91KDa by IPTG induced in Escherichia coli.