【摘要】 构建了一系列含有编码硫氧还蛋白(Trx)和绿色荧光蛋白(GFP)基因的双顺反子重组表达质粒,重点考察双顺反子间紧密相连和重迭的间隔序列。诱导表达后,对表达的GFP进行荧光检测,利用GFP的荧光强度来反映蛋白质的表达水平。结果表明各表达载体所表达的GFP的平均荧光强度差距很大,表明双顺反子间的间隔序列对报告基因表达水平有一定的影响,为实现不同功能基因表达的精确调控奠定了基础。更多还原

【Abstract】 The spacer sequences of polycistron of genome in E.coli,especially the one that the stop codon of the upstream cistron abuts directly the start codon of the downstream one or overlaps was to be analyzed.A series of expression vectors with a bicistron coding green fluorescent protein(GFP) and Thioredoxin(Trx) were constructed in E.coli,and control plasmids pET32a-GFP and pET32a-Trx which could only expresse gfp or trx were also constructed.These plasmids were transformed to BL21(DE3) and induced by IPTG.The results suggested that average fluorescence intensive ratio of pET32a-GFP,pET32a-Trx-GFPR,pET32a-Trx-GFPL,pET32a-Trx-GFPD and pET32a-Trx was 970.35,762.82,12.974,11.810 and 4.4168,respectively.It proved effect of spacer sequences of bicistrons on expression levels of GFP,and also provided a basis for regulation and control of gene expression.更多还原