实验以ｐＢｌｕｅｓｃｒｉｐｔｌｌｋｓ质粒为载体，构建了人白细胞介素６次级克隆ｐＢＩＬ－６。用限制性内切ＢａｍＨＩ和ＥｃｏＲＶ消化ｐＢＩＬ－６，分离并回收了ＩＬ－６ｃＤＮＡ片段，并在其平未端加入了ＢａｍＨＩ接头。实验将带有ＢａｍＨＩ粘性末端的ＩＬ－６全基因片段插入到了逆转录病毒载体ｐＺＬＰＮｅｏＳＶ（Ｘ）１的ＢａｍＨＩ位点上，构建了重组质粒ｐＺＬＰＩＬ－６。经对重组予ＤＮＡ的琼脂糖凝胶电泳、限制住内切酶分析和核酸杂交鉴定，筛选出了含有ＩＬ－６ｃＤＮＡ片段及插入方向正确的ｐＺＬＰＩＬ－６。 The pBluescriptllks plasmid was used as a vector to construct human interleukin 6 secondary clone pBIL-6. The pBIL-6 was digested with restriction endonucleases BamHI and EcoRV, the IL-6 cDNA fragment was isolated and recovered, and a BamHI linker was added at its blunt end. The IL-6 whole gene fragment with BamHI cohesive ends was inserted into the BamHI site of the retroviral vector pZLPNeoSV (X) 1 to construct the recombinant plasmid pZLPIL-6. The recombinant DNA was subjected to agarose gel electrophoresis, restriction endonuclease analysis and nucleic acid hybridization were identified, and the pZLPIL-6 containing the IL-6 cDNA fragment and the correct orientation was screened out.