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.
出处
《西安交通大学学报（医学版）》
CAS
CSCD
北大核心
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)
