Dobrynin V N, Boldyreva E F, Filippov S A, Chuvpilo S A, Korobko V G
Bioorg Khim. 1987 Jan;13(1):119-21.
A plasmid vector (pEK1) coding, in framework of beta-galactosidase gene, for the amino acid sequence (Asp)4Lys which is recognized by bovine enteropeptidase has been constructed. Using this vector and chemically synthesized DNA coding for the [Leu5]-enkephalin, a plasmid (pEK-ENK) has been obtained in which the beta-galactosidase gene is fused, through the enteropeptidase linker, with the gene for [Leu5]enkephalin. The chimeric protein produced by expression of this plasmid has been isolated and then cleaved by the enteropeptidase to give [Leu5]enkephalin with the yield 74%.
已构建了一种质粒载体(pEK1),其在β-半乳糖苷酶基因框架内编码牛肠肽酶可识别的氨基酸序列(Asp)4Lys。利用该载体和化学合成的编码[Leu5]-脑啡肽的DNA,获得了一种质粒(pEK-ENK),其中β-半乳糖苷酶基因通过肠肽酶连接子与[Leu5]脑啡肽基因融合。通过表达该质粒产生的嵌合蛋白已被分离出来,然后用肠肽酶切割,得到[Leu5]脑啡肽,产率为74%。