一种含ELP60基因的载体pRELPN促进大肠杆菌的外源基因的高表达

A pRELPN Vector Containing the ELP60 Gene Promotes the High Expression of Heterologous Genes in E. coli

类弹性蛋白多肽(ELP)为含有人工合成的ELP60基因的表达载体pRELPN,能促使外源基因在大肠杆菌中的高表达。当ELP60在大肠杆菌表达载体p ET28a的多克隆位点被克隆后,其自身的表达低,也不与目的基因构成ELP融合蛋白质,而是促进克隆在ELP60基因后的含起始密码ATG的外源目的基因独立高表达。外源目的基因表达量占宿主蛋白的20%~60%,比用p ET28a载体表达的外源基因表达量高2~10倍。此类表达载体pRELPN适合于表达包括抗体、抗原、酶、重组蛋白质、多肽及ELP融合蛋白质等的外源基因的独立高表达。这些结果表明,pRELPN代表了一种有效的表达载体,有助于解决在原核表达中,所受限的普通载体对外源基因低表达或不表达所导致的不能产业化的问题。

The pRELPN plasmid is an expression vector containing the elastin-like polypeptide（ ELP）gene and can promote the high expression of heterologous genes in E. coli. When ELP60 was cloned in the polyclonal sites of the E. coli expression vector p ET28 a,its own expression was low and the ELP fusion protein was not constructed with the target gene. It also promotes the independent high expression of the heterologous gene containing the initiation codon ATG after the cloning of the ELP60 gene. The expression of exogenous target gene accounted for 20%-60% of host proteins,which were 2-10 times higher than that of the p ET28 a vector. Therefore, pRELPN is suitable for the independent high expression of the heterologous genes,including antibodies,antigens,enzymes,recombinant proteins,polypeptides,as well as ELP fusion proteins. These results indicate that pRELPN represents an efficient vector to solve the problem of low expression or non-expression of heterologous gene in prokaryotic expression for industrialization.