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Use of Interferon-γ Enzyme-linked Immunospot Assay to Characterize Novel T-cell Epitopes of Human Papillomavirus
Published on: March 8, 2012
[Prokaryotic expression of HPV 16 L1 gene and identification of its immune activity]
Dong-feng Hao1, Zheng-hai Ma, Yan-ping Wang
1Key Laboratory of Xinjiang Biological Resources and Gene Engineering, College of Life Sciences and Technology, Xinjiang University, Urumqi 830046, China.
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.
