人白细胞介素18cDNA的克隆及序列测定

Cloning and Sequencing of Human Interleukin 18cDNA

目的获取人ＩＬ－１８ｃＤＮＡ克隆。方法用ＲＴ－ＰＣＲ方法从人外周血单个核细胞中获取人ＩＬ－１８ｃＤＮＡ，将其克隆至天然蛋白表达载体ＰＢＶ２２０中，并进一步亚克隆至融合蛋白表达载体ＰＧＥＸ－４Ｔ－３中，对其进行酶切分析及序列测定。结果测序结果表明克隆至ＰＢＶ２２０中的ＩＬ－１８ｃＤＮＡ包含成熟ＩＬ－１８蛋白的全部编码序列，酶切分析表明一约４７０ｂｐ的基因片段克隆至ＰＧＥＸ－４Ｔ－３中，与理论预计的一致。结论获得了编码人ＩＬ－１８成熟活性蛋白的ｃＤＮＡ克隆。

Objective To obtain the gene encoding human IL 18 mature active protein.Methods The human IL 18cDNA obtained by RT PCR from PBMC was cloned into PBV220 and subcloned into PGEX 4T 3.The recombinant plasmid was characterized with sequencing and restriction enzyme digestion.Results The result of PBV220 IL 18cDNA sequencing showed that the sequence accorded with that published completely, and restriction enzyme degestion suggested that a fragment of about 470 bp was subcloned into PGEX 4T 3.Conclusion The gene encoding full human IL 18 active protein was obtained.