含翻译增强子的表达载体pSC34的构建及初步应用

１９８８年Ｏｌｉｎｓ［１］发现Ｔ７噬菌体基因１０的先导序列（Ｔ７ｇ１０Ｌ）具有明显的促进基因翻译的作用．本文构建了含Ｔ７ｇ１０Ｌ的表达载体ｐＳＣ３４，并尝试表达了几个不同类型的基因．１材料与方法１．１菌株大肠杆菌菌株ＴＡＰ１０６为本室储存．ＴＡＰ１０...

Construction and Application of Expression Vector with Translational Enhancer

In order to improve gene expression of the prokaryotic system, it is critical to enhance the translation efficiency of mRNA. On this basis, one new expression vector pSC34 was constructed by inserting the translation enhancing sequence of T7g10L into the up stream of SD sequence within a two cistron vector pCC11 constructed also. In the same time the first cistron region of the previous vector was cut off. The properties of the vector is that it is relatively small without cI857 gene so convenient to be operated; its host strain is E.coli TAP106 with the genome containing cI857; its promoter P L is heat inducing and can be controlled stringently; the two initial ones of multiple cloning sites, Nsi Ⅰ and Sph Ⅰ, can serve for the translation initiation of target gene with or without start codon ATG respectively. By use of the vector, several genes had been overexpressed high efficiently, such as E.coli gene trxA , human protein disulfide isomerase gene, human IL 3 gene and synthetic human IFN gamma gene. It has been confirmed primarily that the vector can be used to express foreign genes with high efficiency and well generality.