人白细胞介素6逆转录病毒表达载体的构建  

CONSTRUCTION OF RETROVIRAL VECTOR FOR EX-0PREXXING HUMAN INTERILUKIN-6

作　　者：黎诚耀[1] 费恩阁[1] 王世若[1] 韩文瑜[1] 高荣凯[1] 

机构地区：[1]解放军农牧大学微生物学教研室

出　　处：《中国免疫学杂志》1994年第2期81-83,共3页Chinese Journal of Immunology

摘　　要：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 secondary cloning vector pBIL-6 carrying human interleukin-6 cDNA was constructed with pBluescript 11ks plasmid.IL-6 cDNAfragments were isolated from pbIL-6degested by EcoR V and BamHI,and BamHI,linkers were ligated to the blunt ends of IL-6cDNA fragments.The experiments constructed the recombinant plasmid pZIPIL-6 by insert-ing IL-6 cDNA with BamHI cohesive termini into the BamHI site of the tetroviral vectorpZIPNeoSV(X)1.Screening and identifying for the recombinant plasmids by agrose geleledtrophorcsis,restriction enzyme analysis and nucleic acid hybridzation,the results evi-denced that pZIPIL-6 contained the correctly inserted IL-6 cDNA fragment.This study pro-vides a fundament for using recombinant retroviru-mediated IL-6 gene transfer and therapy.

关 键 词：白细胞介素6 病毒载体 基因 

分 类 号：R392.12[医药卫生—免疫学]