Carboxyl terminus truncated HPV58 virus L1 protein expressed with baculovirus system and its bioactivity.
- Author:
Wen-Sheng LI
1
;
Jin ZHENG
;
Hong-Li LIU
;
Hong-Wei CHEN
;
Jun YANG
;
Yi-Li WANG
;
Lü-Sheng SI
Author Information
1. Institute for Cancer Research School of Life Science & Technology, Xi'an Jiaotong University, Xi'an 710061, China.
- Publication Type:Journal Article
- MeSH:
Animals;
Baculoviridae;
genetics;
Blotting, Western;
Cloning, Molecular;
Humans;
Mice;
Mice, Inbred C57BL;
Papillomaviridae;
genetics;
Papillomavirus Vaccines;
immunology;
Recombinant Proteins;
biosynthesis;
isolation & purification;
Vaccines, Synthetic;
immunology;
Viral Proteins;
biosynthesis;
genetics;
Virion;
immunology
- From:
Chinese Journal of Biotechnology
2004;20(4):536-539
- CountryChina
- Language:Chinese
-
Abstract:
To prepare carboxyl terminus truncated human papillomavirus type 58L1 protein, and study on its in vitro bioactivity. PCR was used to amplify carboxyl terminus truncated HPV 58L1 gene, the product was inserted into the PUCMT cloning vector, preparing recombinant PFastBacHTb containing carboxyl terminus truncated HPV58L1 gene. Further more, the recombinant plasmid PfastbacHTb was used to transform DH10Bac cells, constructing recombinant Baculovirus, then the recombinant virus was successfully used to infect Sf-9 insect cells. After incubating at 27 degrees C for 72 hours, the infected cells were collected and total cellular proteins were extracted. The target protein with MW 58KD was revealed by SDS-PAGE and confirmed by Western blot. The interested protein was purified by ProBond purification system. The purified interested protein was identified to self-assemble into VLPs by Transmission electron microscope, and induce murine erythrocyte hemagglutination, indicating that the given proteins had the conformation of VLPs, collecting, HPV58L1 proteins with carboxyl terminus truncation could be efficiently expressed in baculovirus Sf-9 cells expression system, it has identical in vitro bioactivity to the wild type HPV58L1, The present study is fundmental for preparing HPV58L1 prophylactic vaccine.
