组织型纤溶酶原激活剂cDNA克隆及其乳腺特异性表达载体的构建

Cloning of tPA cDNA and construction of mammary-specific expression vector

目的 :克隆组织型纤溶酶原激活剂 (tissue typeplasminogenactivator,tPA)基因及构建其乳腺特异性表达载体。方法 :以PCR技术克隆 2 .1kbtPAcDNA片段 ;应用体外连接技术 ,连接乳腺特异性表达载体pWA和 2 .1kbtPAcDNA片段 ;转化连接产物 ,以内切酶酶切、琼脂糖电泳及插入片段序列分析鉴定阳性克隆。结果 :琼脂糖电泳显示 ,2 .1kbtPAcDNA片段克隆成功 ;内切酶酶切片段与预期片段长度一致 ;插入片段序列与文献报道一致。结论

Aim: To clone tissue type plasminogen activator (tPA) cDNA and construct the mammary specific expression vector. Methods: The 2.1 kb tPA cDNA fragment was cloned by PCR. The tPA cDNA and pWA plasmid were ligated in vitro to form vector pWTA. The pWTA plasmid was identified by endonuclease digestion and the inserted fragment sequencing. Results: By agarose electrophoresis, the tPA cDNA was demonstrated. The endonuclease digestion map of pWTA was consistent with the fragment length in anticipation. Sequencing of the inserted fragment was the same to what reported. Conclusion: The tPA gene cloning and mammary specific expression vector were successfully established.