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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 ,表达产物可与相应的抗血清发生阳性反应
OBJECTIVE: To obtain a sufficient amount of human papillomavirus type 16 (HPV16) L1 fusion protein and a gene recombinant containing the HPV 16 L1 ORF sequence. Methods: The HPV16 L1 gene fragment was amplified by PCR. The fragment was inserted into prokaryotic expression vector pGEMEX 1, and the corresponding recombinant plasmid pGEMEX L1 was constructed. The plasmid was transformed into E. coli JM109 (DE3) Western blotting of bacterial proteins was performed after induction of expression. Results: The recombinant HPV16 L1 gene was constructed successfully. The fusion protein expressed in Escherichia coli produced by the cloned target gene fragment has the antigenicity of anti-HPV16 L1-positive serum in Western blotting assay. Conclusion: The HPV16 L1 gene fragment can be expressed efficiently in E. coli, and the expression product can react with the corresponding antiserum