孔石莼凝集素蛋白基因的克隆与表达

Cloning and Expression of Lectin Gene of Ulva pertusa

根据孔石莼(Ulva pertusa)凝集素(Lectin)蛋白cDNA全长序列(GenBank登录号:AY433960)设计引物,以其总DNA为模板,采用PCR技术扩增蛋白DNA序列,经克隆、测序获得基因序列。结果表明,孔石莼凝集素蛋白(UPL)基因序列长约为670 bp,含有一个大小为56 bp的内含子。此外,设计带酶切位点的引物,以UPL-cDNA为模板,扩增其开放阅读框,并与表达载体pGEX-2T连接,构建原核表达载体pG2T-UPL,并在大肠杆菌BL21(DE3)中成功表达大小约为47 kD的目的蛋白。

Based on the known cDNA sequence of lectin protein gene of U. pertusa （GenBank accession number： AY433960）, a pair of primers were designed and used to clone, and sequence the lectin protein gene with PCR technique by using the genomic DNA of U. pertusa as the template. The results showed that the lectin protein gene consisted of 670 bp, with a 56 bp intron. A pair of primers with restriction enzyme sites were designed to amplify the ORF using the genomic cDNA of U. pertusa as the template. The ORF was inserted into the expression vector pGEX-2T and a protein with molecular weight of 47 kD was successfully expressed in Escherichia coli BL21 （DE3）.