PAI-1基因的克隆

Cloning of PAI-1 gene

目的构建PAI-1真核细胞表达质粒。方法根据GenBank中PAI-1的序列，利用RT—PCR从人卵巢癌组织中进行扩增PAI-1基因全长片段，将其克隆人真核表达质粒pcDNA3．1／myc—his（-）B，建立PAI-1的表达质粒pcDNA3．1-PAI-1，并对克隆的全长片段进行DNA序列测定。结果经与GenBank分布的序列进行分析比较证实，构建的真核表达重组质粒pcDNA3．1-PAI-1含有PAI-1全长cDNA编码序列。结论PAI-1能成功地从人卵巢癌组织中克隆，构建了真核细胞表达质粒pcDNA3．1-PAI-1，测序表明其携带有PAI-1的全长序列，为进一步研究PAI-1在卵巢癌侵袭转移中的作用奠定了基础。

Objective：To construct the expression plasmid carrying PAI-1 gene. Method：The cDNA encoding the PAI-1 was amplified by using RT -PCR method from the human ovarian tumor tissue according to its GenBank sequence. Being isolated and Inserted PAI-1 gene into pcDNA3. 1/myc -his（-）B an expression vector containing PAI-1 gene, pcDNA3. 1-PAI-1 was constructed by using recombinant DNA techniques. The recombinant was analyzed and identified by restriction enzyme,PCR and sequencing. Results： The PAI-1 gene was successfully cloned into pcDNA3. 1/myc-his （-）B. The sequence of the PAI-1 showed the same as its sequence in the GenBank. Conclusion： The construction of an expression system of the PAI -1 establishes the foundation for further research on the role of the PAI-1 in the invasion and metastasis of the ovarian cancer.