HPV16L1ORF的基因克隆及在大肠杆菌中的表达

目的获得足够量的人乳头瘤病毒16型(HPV16)L1融合蛋白及含HPV16L1ORF序列的基因重组体。方法通过PCR扩增获得HPV16L1基因片段，将此片段插入原核细胞表达载体pGEMEX-1，构建相应的重组表达质粒pGEMEX-L1，将此质粒转化大肠杆菌JMl09(DE3)，得到的转化子经IPTG诱导表达后进行菌体蛋白的Western blotting免疫印迹分析。结果HPV16L1基因重组体构建成功，克隆的目的基因片段在大肠杆菌中产生的融合表达产物在Western blotting检测中具有和抗HPV16L1阳性血清反应的抗原性。结论HPV16L1基因片段可在大肠杆菌中得到有效表达，表达产物可与相应的抗血清发生阳性反应。

CLONING AND EXPRESSION OF HUMAN PAPILLOMAVIRUS TYPE 16 L1 ORF IN E.COLI

ObjectiveTo obtain enough HPV16 L1 fused protein and recombinant plasmid containing HPV16 L1 gene. MethodsHPV16 L1 gene was amplified with PCR. The HPV16 L1 gene was first inserted into the pGEM-T easy vector and then was inserted into the expressing vector pGEMEX-1, to construct a recombiant expressing plasmid,which was transformed into the E.coli Jm109(DE3) and the IPTG induced transformant was analyzed with Western blotting. Results The induced expressing product was shown to have antigenic reactivity to HPV16 L1 antibody positive sera. Conclusion The HPV16 L1 gene was expressed effectively in E.coli.