HPV16L1 ORF的基因克隆及在大肠杆菌中的表达

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

目的 :获得足够量的人乳头瘤病毒 16型 (HPV16)L1融合蛋白及含HPV 16L1ORF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JM 10 9(DE3) ,得到的转化子经IPTG诱导表达后进行菌体蛋白的Westernblotting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Westernblotting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 。

Objective:To obtain enough HPV16 L1 fused protein and recombinant plasmid containing HPV16 L1 gene.Methods:HPV16 L1 gene was amplified with PCR.The HPV16 L1 gene was first inserted into the pGEMT easy vector and then was inserted into the expressing vector pGEMEX1,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.