Carboxyl terminus truncated human papillomavirus type 58 L1 protein maintains its bioactivity and ability to form virus-like particles.
To prepare carboxyl terminus truncated human papillomavirus type 58 L1 (HPV58L1) protein and evaluate its ability to form virus-like particles, the baculovirus and Sf-9 insect cells was used to express HPV58L1 protein, and pFastBac-Htb containing HPV58L1 gene sequence of carboxyl terminus truncation was generated. Then Sf-9 cells were infected with recombinant baculovirus. After being cultured, the post-infected cells expressing--HPV58L1 protein were harvested and analyzed by SDS-PAGE and Western blot. The ProBond purification system was used for protein purification. The bio-activity of purified protein was identified by mouse erythrocyte hemagglutination assay, and the VLP formation was examined with transmission electron microscope. Our results showed that the recombinant baculovirus was generated and the Sf-9 cells was infected with the recombinant baculovirus, and after collecting, total cellular proteins were extracted. Truncated HPV58L1 protein with MW 58KD was revealed by SDS-PAGE and confirmed by Western blot. The purified L1 proteins under native condition could cause mouse erythrocytes to agglutinate and form VLP. It is concluded that HPV58L1 protein with carboxyl terminus truncation could be efficiently expressed. In baculovirus Sf-9 cells expression system, the purified protein could self-assemble into virions in vitro, and induce agglutination of mouse erythrocytes, indicating that carboxyl terminus truncation does not interfere with the bioactivity of HPV58L1 protein.
Li
Wensheng
W
The Key Laboratory of Biomedical Information Engineering of Ministry of Education, Institute for Cancer Research, School of Life Science and Technology, Xian Jiaotong University, Xian 710061, China.
Liu
Hongli
H
Zheng
Jin
J
Chen
Hongwei
H
Yang
Jun
J
Wang
Lixiu
L
Yan
Xiaofei
X
Wang
Yili
Y
Si
Lüsheng
L
D016367:Baculoviridae / Q000235:genetics / Q000378:metabolism; D036022:Capsid Proteins; D002245:Carbon Dioxide; D006801:Humans; D007181:Inclusion Bodies, Viral / Q000648:ultrastructure; D009856:Oncogene Proteins, Viral / Q000096:biosynthesis* / Q000235:genetics / Q000276:immunology; D027383:Papillomaviridae / Q000502:physiology*; D011994:Recombinant Proteins / Q000096:biosynthesis; D014771:Virion / Q000254:growth & development* / Q000502:physiology; D019065:Virus Assembly
D036022:Capsid Proteins; C114580:HPV L1 protein, Human papillomavirus; D009856:Oncogene Proteins, Viral; D011994:Recombinant Proteins; D002245:Carbon Dioxide; C041069:carboxyl radical