pcDNA3.1- Bcl-2真核表达载体构建

目的 克隆大鼠Bcl-2基因（B cell lymphoma）全长cDNA，构建含大鼠Bcl-2基因的重组真核表达质粒。方法 采用RT-PCR的方法从大鼠脾脏组织中扩增全长Bcl-2cDNA,将扩增产物克隆至pMD18-T载体, 测序证实后，再克隆至真核表达载体pcDNA3.1(+),构建了pcDNA3.1- Bcl-2重组质粒。结果: RT-PCR获得长度为720 bp的阳性产物,经T载体和pcDNA3. 1(+)真核表达载体克隆、酶切鉴定及序列分析后,证实真核表达质粒pcDNA3.1- Bcl-2构建成功。结论 成功克隆了Bcl-2基因，并构建了pcDNA3.1- Bcl-2重组质粒,为进一步研究Bcl-2基因的功能及应用Bcl-2进行基因治疗奠定基础。

Construction of pcDNA3.1-Bcl-2 expression vector

Objective　To construct the pcDNA3.1-Bcl-2 expression vector after cloning the bcl-2 gene. Methods　 Bcl-2-cDNA was obtained and identified by RT-PCR from kidney tissue of a Wistar rat,and was cloned into pMD18-T vector.The Bcl-2 gene was inserted into the vector pcDNA3.1(+) after sequencing ..Results　A 720 bp fragment was obtained by RT-PCR. After double enzyme digestion, the Bcl-2 gene was inserted into pcDNA3. 1(+) vector.Conclusion　 Bcl-2-cDNA was cloned and was constructed into eukaryotic expression vector pcDNA3.1- Bcl-2 successfully.