4-1BBL胞外区cDNA的克隆及鉴定

Cloning and Characterization of cDNA at Extracellular Domain of 4-1 BBL

目的 研究共刺激分子4-1BBL对T淋巴细胞的作用,并进一步研究肿瘤的免疫治疗。方法 按照4-1BBL的基因序列,设计合成了可扩增4-1BBL胞外区的引物序列,用RT-PCR方法从急性B淋巴白血病Raji细胞系细胞中提取总RNA,扩增出4-1BBL胞外区片段,并克隆到载体中,经酶切鉴定后再进行序列分析。结果 所克隆的目的片段经EcoRI和XhoI双酶切和序列分析证明正确。结论 已成功地钓取4-1BBL胞外区基因并克隆到pMD-18载体上。

Objective To study the effect of 4-1 BBL on T lymphocyte and the role of it in immuno-therapy of tumors.Methods Design and synthesize primers according to the gene sequence of 4-1 BBL, extract total RNA from acute B lymphocyte leukemia Raji cell line by RT-PCR, amplify the cDNA fragment at extracellular domain of 4-1 BBL and clone into expression vector, digest with restriction endonuclease and sequence. Results Digestion with EcoRI and Xhol and sequencing proved correct sequence of cloned goal .fragment. Conclusion The cDNA at extracellular domain of 4-1 BBL was successfully extracted and cloned into pMD-18 vector.