Lp-1643蛋白单一结构域的黏附功能研究

Adhesion capacity of the single domain of Lp-1643 protein

从植物乳杆菌KLDS 1.0320中克隆出Lp-1643蛋白N端的第一个结构域基因,与表达载体pET30a连接后成功构建重组质粒pET30a/N1。重组质粒转化大肠杆菌BL21后,以IPTG进行诱导,重组蛋白以可溶形式成功获得表达。通过亲和色谱技术用HisTrap FF柱对重组蛋白进行了分离纯化。以BSA作为对照,研究了重组蛋白His-N1对KLDS 1.0320菌株黏附Caco-2细胞的影响,发现用该重组蛋白预处理Caco-2细胞之后,其上黏附的KLDS 1.0320数量显著减少(P<0.05)。这说明Lp-1643蛋白N端的第一个结构域具有黏附Caco-2细胞的功能。

The gene coding for the first domain of the N terminus of Lp_ 1643 protein was amplified from KLDS 1.0320 genome. An expression vector designated pET30a/N1 was constructed. The recombinant protein was highly expressed in the strain E. co/i BL21 induced by IPTG. Affinity chromatography with HisTrap FF column was used to separate and purify the recombinant protein of His-N1. Influences of the recombinant protein His-N1 on adhesion capacity of strain KLDS 1.0320 to Caco-2 cells were studied with BSA as a negative control. It was found that the numbers of KLDS 1.0320 adhering to Caco-2 cells pretreated by His-N1 dramatically decreased. 31fis result indicates the single domain of Lp_ 1643 protein can adhere to Caco-2 cells.