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目的:在大肠杆菌中表达人乳头瘤病毒16型(HPV16)主要衣壳蛋白L1,并鉴定其免疫反应性。方法:将HPV16L1基因克隆入原核表达载体pThioHisC中,构建重组表达载体。以重组载体分别转化大肠杆菌Top10和DH5α,在IPTG诱导下表达外源基因,用SDSPAGE和Westernblot对表达产物进行鉴定和分析。结果:构建了HPV16L1基因的原核表达质粒pThioHisC/HPV16L1,并在大肠杆菌中表达出相对分子质量(Mr)约为70800的蛋白。表达的蛋白能与抗HPV16L1抗体发生特异性反应。结论:在原核细胞中成功地表达HPV16L1基因,为HPV16L1疫苗的研制提供了必要的基础。
OBJECTIVE: To express human papillomavirus type 16 (HPV16) major capsid protein L1 in Escherichia coli and identify its immunoreactivity. Methods: The HPV16 L1 gene was cloned into the prokaryotic expression vector pThioHisC to construct a recombinant expression vector. The recombinant vectors were respectively transformed into E. coli Top10 and DH5α, and induced by IPTG to express foreign genes. The expression products were identified and analyzed by SDSPAGE and Western blotting. Results: The prokaryotic expression plasmid pThioHisC / HPV16L1 of HPV16 L1 gene was constructed and the protein with relative molecular mass (Mr) of about 70800 was expressed in E. coli. The expressed protein can specifically react with anti-HPV16 L1 antibody. Conclusion: The successful expression of HPV16 L1 gene in prokaryotic cells provided the necessary foundation for the development of HPV16 L1 vaccine.