人IL-16的原核表达及活性检测

The Prokaryotic Expression and Activity Assay of Recombinant Human IL-16

用RT PCR方法将人IL 16基因的活性区段 (783 1187bp )克隆至pBluescriptSK (+ )载体 ,测序验证后 ,克隆至原核表达载体pT7 7His ,并在大肠杆菌中进行了高效表达。表达产物为可溶性蛋白 ,分子量约为 18kD ,通过Ni+离子亲和层析一次性纯化。Westernblot检测证实其具IL 16免疫原性 ,FACS检测证实该rhIL 16具促CD4+细胞增殖活性。

The human IL-16 biological active fragment (783-1187bp) was obtained by RT-PCR from PBMCs,and was cloned into pBluescript SK(+) vector After the sequence was confirmed, it was cloned into the prokaryotic expression vector pT7-7His The expressed product was soluble,MW 18kD,which was purified by Ni + metal chelate affinity chromatography Its antigenic activity was assayed by Western blotting,and the ability to induce the proliferation of CD4 + cell was confirmed by FACS assay