NT4-p53(N15)-Ant融合基因的克隆和鉴定  

Construction and identification of fusion gene of NT4-p53(N15)-Ant

作　　者：宋丽萍[1] 李跃萍[1] 邱曙东[2] 杨广笑 王全颖 

机构地区：[1]西安交通大学医学院第一附属医院放疗科,陕西西安710061 [2]西安交通大学医学院人体解剖与组织胚胎学系,陕西西安710061 [3]西安华广生物工程公司,陕西西安710025

出　　处：《西安交通大学学报（医学版）》2006年第4期333-336,共4页Journal of Xi’an Jiaotong University（Medical Sciences）

基　　金：国家自然科学基金资助项目(No.30471942);陕西省科技攻关项目(No.2004k11-G3)

摘　　要：目的构建NT4-p53(N15)-Ant融合基因表达盒并进行序列分析。方法应用互为模板的引物PCR技术及T载体克隆法克隆p53(N15)-Ant基因,筛选阳性克隆、酶切鉴定并测序。扩增阳性重组质粒后限制性内切酶切取p53(N15)-Ant片段连入pBV220/NT4质粒。结果克隆了p53(N15)-Ant基因,经酶切及测序证实结果正确;重组质粒pBV220/NT4p53(N15)Ant经限制性内切酶及琼脂糖凝胶电泳,结果显示酶切片段大小和理论值一致。结论通过分子克隆体外重组技术成功制备了含有NT4-p53(N15)-Ant表达盒的pBV220质粒,为进一步开展肿瘤的基因治疗奠定了基础。Objective To construct the expression box of fusion gene NT4-p53（N15）-Ant. Methods The gene of p53 （N15）-Ant was obtained by PCR of two primers templating with each other and T-vector cloning method. The positive clone was identified and analyzed by the restriction enzymes and sequencing respectively. After digested with restriction enzyme, the interest gene of p53（N15）-Ant was subcloned into the plasmid pBV220/ NT4. Results The gene of p53（N15）-Ant was confirmed by the digestion of restriction enzyme and sequencing. The recombinant plasmid pBV220/NT4p53 （N15）Ant was identified by the digestion of restriction enzyme and agarose gel electrophoresis and the results conformed theoretical values. Conclusion The plasmid pBV220 containing the expression box of NT4-p53 （N15）-Ant was successfully constructed by molecular cloning and recombination techniques in vitro, which will guide further study on gene therapy of cancer.

关 键 词：p53(N15)-Ant NT4信号肽 基因克隆 肿瘤 

分 类 号：R512.3[医药卫生—内科学] R733.7[医药卫生—临床医学]