摘要
用限制酶BamHI切割质粒pDO432并经琼脂糖凝胶电泳分离,将所得荧光素酶编码序列作为插入片段。用Hind Ⅲ和BamH Ⅰ切割pSVK100去除galk基因及部分tsplice序列作为载体,构建了一对其启动区和荧光素酶编码序列位向相反的质粒,pSV-Luc19及pSV-Luc20。正位向的pSV-Luc20能在爪蟾卵母细胞中转录,翻译并产生活性酶蛋白。结果表明:荧光素酶基因和爪蟾卵母细胞可以构成一个监测启动子活性的有效系统。
A couple of fusion plasmids, pSV-Luc20 and pSV-Luc19,were cons-tructed by using the larger.BamHI restricted fragment of plasmid pSVK100 and the smaller HindIII-BamHI restricted fragment of plasmid pDO432.They have different fusion orientations between the coding sequenceof the luciferase and SV-40 promoter, they are sense and anti-senseplasmids. The results indicated that pSV-Luc20 could be expressed and lucife-rase with high activity was formed when the plasmid was introducedinto Xenophus oocytes by micro-injection.Therefore, the luciferase gene-Xenophus oocyte is an excellent system for monitoring the promoter ac-tivity.
出处
《生物工程学报》
CAS
CSCD
北大核心
1991年第4期318-322,共5页
Chinese Journal of Biotechnology
关键词
荧光素酶
爪蟾
卵母细胞
基因表达
Luciferase
oocyte of Xenophus laevis
gene expression
