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ISSN
1672-9145
Acta Biochim Biophys Sin
2005, 37(3): 153–158
CN 31-1940/Q
Expression of Human Papillomavirus Type 16 L1 Protein in Transgenic
Tobacco Plants
Hong-Li LIU1,2, Wen-Sheng LI1, Ting LEI1,
Jing ZHENG1, Zheng ZHANG3, Xiao-Fei YAN1,
Zhe-Zhi WANG3, Yi-Li WANG1, and Lü-Sheng SI1*
1Institute of Cancer Research, School of Life
Science & Technology, Xi’an Jiaotong University, Xi’an 710061, China;
2First Hospital of Xi’an Jiaotong
University, Xi’an 710061, China;
3Shaanxi Normal University, Xi’an 710062, China
Abstract To develop a plant expression system for the production of the human
papillomavirus type 16 (HPV16) vaccine, we investigated whether the HPV16 L1
protein can be expressed in tobacco plants and whether it can be used as the
cheapest form of edible vaccine. The HPV16 L1 coding sequence was amplified by
PCR using specific primers from the plasmid pGEM-T-HPV16 containing the
template sequence, and subcloned into the intermediate vector pUCmT and binary
vector pBI121 consecutively to obtain the plant expression plasmid pBI-L1. The
T-DNA regions of the pBI-L1 binary vector contained the constitutive
Cauliflower mosaic virus (CaMV) 35S promoter and the neomycin
phosphotransferase npt II gene, which allowed the selection of
transformed plants using kanamycin. The tobacco plants were transformed by
co-cultivating them, using the leaf disc method, with Agrobacterium
tumefaciens LBA4404, which harbored the plant expression plasmid. The
regenerated transgenic tobacco plants were selected using kanamycin, and
confirmed by PCR. The results of the Southern blot assay also showed that the
HPV16 L1 gene was integrated stably into the genome of the transformed
tobacco plants. The Western blot analysis showed that the transformed tobacco
leaves could express the HPV16 L1 protein. Furthermore, it was demonstrated by
ELISA assay that the expressed protein accounted for 0.034%0.076% of the total
soluble leaf protein, was able to form 55 nm virus-like particles compatible
with HPV virus-like particle (VLP), and induced mouse erythrocyte
hemagglutination in vitro. The present results indicate that the HPV16
L1 protein can be expressed in transgenic tobacco plants and the expressed
protein possesses the natural features of the HPV16 L1 protein, implying that
the HPV16 L1 transgenic plants can be potentially used as an edible
vaccine.
Key words human papillomavirus
(HPV); virus-like particle; transgenic tobacco; plant vaccine; Agrobacterium
tumefaciens; L1 major capsid protein
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Received: October 9, 2004 Accepted: January 28,
2005
This work was supported by a grant from the National Natural Science
Foundation of China (No. 30070848)
*Corresponding author: Tel, 86-29-82655191; Fax, 86-29-82655499;
E-mail, [email protected]