[人乳头瘤病毒16型L1基因的原核表达及其免疫活性鉴定]
[Prokaryotic expression of HPV 16 L1 gene and identification of its immune activity].
作者信息
Hao Dong-feng, Ma Zheng-hai, Wang Yan-ping, Zhang Fu-chun
机构信息
Key Laboratory of Xinjiang Biological Resources and Gene Engineering, College of Life Sciences and Technology, Xinjiang University, Urumqi 830046, China.
出版信息
Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi. 2005 Jul;21(4):428-31.
AIM
To express the major capsid protein of human papillomavirus type 16 L1(HPV 16 L1) in E.coli and identify its immune activity.
METHODS
The L1 gene of HPV 16 was cloned into the expression vector pThioHisC. The recombinant expression vector was transformed into E.coli, and the HisC-L1 protein was expressed under IPTG induction. The fusion protein was characterized by SDS-PAGE and Western blot.
RESULTS
The HPV16-L1 gene in plasmid pThioHisC was expressed in E.coli as a fusion protein with M(r) about 70,800. The fusion protein reacted specifically with antibodies against HPV16-L1.
CONCLUSION
The HPV-16 L1 gene was expressed successfully in E.coli, which provides necessary basis for preparing HPV-16 L1 vaccine in human.
目的
在大肠杆菌中表达人乳头瘤病毒16型主要衣壳蛋白L1（HPV 16 L1）并鉴定其免疫活性。
方法
将HPV 16的L1基因克隆到表达载体pThioHisC中。将重组表达载体转化到大肠杆菌中，在异丙基-β-D-硫代半乳糖苷（IPTG）诱导下表达HisC-L1蛋白。通过十二烷基硫酸钠-聚丙烯酰胺凝胶电泳（SDS-PAGE）和蛋白质免疫印迹法（Western blot）对融合蛋白进行鉴定。
结果
质粒pThioHisC中的HPV16-L1基因在大肠杆菌中表达为分子量约为70,800的融合蛋白。该融合蛋白与抗HPV16-L1抗体发生特异性反应。
结论
HPV-16 L1基因在大肠杆菌中成功表达，为在人体内制备HPV-16 L1疫苗提供了必要依据。
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