HL-60细胞中IL6基因的克隆及鉴定

目的：为探索性构建重组人白细胞介素６－绿脓杆菌外毒素融合蛋白（ＩＬ６－ＰＥ４０）以选择性杀伤高表达ＩＬ６受体（ＩＬ６Ｒ）的白血病细胞，本研究试图从人急性早幼粒细胞白血病细胞株（ＨＬ－６０）中克隆Ｎ－末端缺失２４个氨基酸的人ＩＬ６基因（ＩＬ６ｃＤＮＡ）并构建含此基因的重组质粒。方法：根据ＩＬ６基因序列设计合成可扩增ＩＬ６ｃＤＮＡ的特异性引物；利用基因重组技术，构建含ＩＬ６基因的重组质粒ｐＵＣ－ＩＬ６。结果与结论：本文首次从ＨＬ－６０中扩增出预期４８０ｂｐ的ＩＬ６ｃＤＮＡ，将其克隆至ｐＵＣ１８质粒中，命名为ｐＵＣ－ＩＬ６，并经ＥｃｏＲＩ／ＢａｍＨＩ双酶切电泳及序列分析鉴定加以确证，为进一步构建重组人ＩＬ６－ＰＥ４０奠定了坚实基础。

Cloning of IL6 gene from human promyelocytic leukemia cell line

AIM:In order to construct recombinant interleukin 6-Pseudomonas exotoxin fusion protein (IL6-PE40) to selecting kill leukemia cells expressing high levels of IL6 receptors, we tried to clone IL6 gene from human promyelocytic leukemia cell line (HL-60) and construct the recombinant plasmid encoding IL6.METHODS:Two pairs of primes for IL6 were designed and synthesized according to the sequences of human IL6 gene derived from gene bank. A recombinant plasmid pUC-IL6 coding for IL6 was constructed by recombinant gene technique.RESULTS and CONCLUSION:The recombinant plasmid pUC-IL6 was constructed by cloning 480 bp IL6 cDNA fragment into pUC18 plasmid, confirmed by DNA sequencing and EcoRI/BamHI restriction enzyme digesting. The recombinant plasmid pUC-IL6 can be used to construct recombinant interleukin 6-Pseudomonas exotoxin fusion protein (IL6-PE40).