抗人小细胞肺癌T细胞受体嵌合基因的克隆

CLONING OF THE CHIMERIC GENE OF T CELL RECEPTOR AGAINST HUMAN SMALL CELL LUNG CANCER

目的克隆人小细胞肺癌抗体／Ｔ细胞受体嵌合基因。方法应用聚合酶链反应（ＰＣＲ）技术将人小细胞肺癌单克隆抗体的变区ＶＨ、ＶＫ与Ｔ细胞受体（ＴＣＲ）恒区Ｃα、Ｃβ进行拼接，构建成ＶＨＣβ、ＶＫＣα嵌合ＴＣＲ，并分别克隆至ｐＭＡＭ、ｐＸＴ１真核表达载体。结果应用ＰＣＲ中重叠延伸剪接（ＳＯＥ）方法，能简化基因拼接步骤；该嵌合基因的克隆有助于对肿瘤免疫治疗新途径的探索。

PURPOSE To clone SCLC antibody/T cell receptor chimeric gene.METHODS We constructed the chimeric T cell receptor genes( VHCβ,VKCα)composed of the variable domains (VH,VK) from MAB and constant domains (Cα,Cβ) from TCR by the method of PCR. Then,the two chimeric genes are cloned into eukaryotic expression vectors: pMAM, pXT1.RESULTS We cloned the chimeric gene of T cell receptor against human small cell lung cancer successfully and identified it by enzyme restriction analysis. CONCLUSIONS The results demonstrate that it simplifies gene recombination by SOE method and indicate that the chimeric gene may be helpful for new cancer immunotherpy research.