重组人IL-4 cDNA及纯化蛋白的鉴定

IDENTIFICATION OF RECOMBINANT HUMAN INTERLEUKIN-4 cDNA AND PURIFIED RECOMBINANT HUMAN INTERLEUKIN-4 PROTEIN

构建了重组人白细胞介素4(rhIL—4)表达质粒pBV220/rhIL—4a,并在E.coli中高效表达。经核苷酸序列分析,测出rhIL—4cDNA全序列与国外发表的hIL—4cDNA序列完全相同。其纯化蛋白产品经分子量测定,Western印迹分析,等电点分析,N—端氨基酸序列测定,比活性等测定证明,均符合rhIL—4。

A hIL-4 expression plasmid pBV220/hIL-4a was constructed and effectively expressed in E. colt. The DNA sequence encoding mature hIL-4 was determined and displayed to be identical with that already published. The purified rhIL-4 protein was analyzed by means of SDS-PAGA, Western blotting,isoelectric point(PI) determinaltion ,N-termi-nal amino acid sequencing and specific activity. The results showed that the purified protein was identical with recombi-nant hIL-4 and exhibited high level of bioactivity. Thus, we have developed a useful strategy for the expression and pruification of rhIL-4.