人乳头瘤病毒16 E7 (HPV16E7)逆转录病毒载体的构建

目的 构建人乳头瘤病毒16型E7基因片段（HPV16E7）的逆转病毒载体。方法 PCR扩增HPV16cDNA全长的E7片段，并用酶切，连接等方法将HPV16E7亚克隆到逆转录病毒载体pLXSN上，然后再用酶切，测序进行鉴定，通过Genbank的数据库分析软件对PV16E7进行同源性分析。结果 经酶切分析，测序证明，从HPV16cDNA克隆的300bp的HPV16E7与逆转录病毒载体发生了基因重组，HPV16E7基因片段与数据库中的原HPV16cDNA的E7有高度的同源怀。结论 构建了含HPV16E7的逆转录病毒载体，为人关节软骨细胞的基因转染打下了基础。

Construction of retroviral vector encoding for human papillomavirus type 16 E7 protein (HPV16E7)

Objective To construct a retroviral vector of human papillomavirus type 16 E7(HPV16E7). Methods The E7 fragment of HPV16 cDNA was amplified with PCR and inserted into the retroviral vector pLXSN with endonuclease digestion and ligation methods, then the recombinant vector was determined with enzyme digestion and sequencing. The homologous features between the obtained HPV16E7 sequence and original HPV16 cDNA E7 fragment was analyzed with the Genbank data analysis software. Results The 300 bp fragment of HPV16E7 cloned from HPV16 cDNA was transferred into the genome of retroviral vector in success. The obtained HPV16E7 showed highly homology with the original HPV16 cDNA in Genbank. Conclusion Successfully constructed retroviral vector encoding for HPV16E7 establishes important basis for gene transfection of human articular chondrocytes.