[Optimized codon usage enhances the expression and immunogenicity of DNA vaccine encoding the HPV 6b E7 gene].
OBJECTIVE
To analyze the influence of optimal codon usage on the expression levels and immunogenicity of DNA vaccines, encoding the human papillomavirus type 6b (HPV 6b) E7 gene.


METHODS
The full length E7 gene of HPV 6b was modified to substitute human preferred codon for rarely used codon, and three mutations were introduced into the pRB binding site of HPV 6b E7 to eliminate its transformation potential. The codon optimized and mutated E7 gene (hu-mE7) were cloned into the Kpn I and EcoR I site of the pcDNA3 mammalian expression vector, the in vitro expression of the hu-mE7 gene and the immunogenicity of hu-mE7 DNA vaccine were compared with the wt-E7gene.


RESULTS
The in vitro expression of pcDNA3-hu-mE7 was much higher than the classical wt-E7 plasmid in monkey COS-1 cell line. Mice immunized intramuscularly with the pcDNA3-hu-mE7 showed that the codon modified E7 gene induced a stronger IFN-gamma ratios than the wt-E7 gene.


CONCLUSIONS
These results suggest that the optimized codon usage contributes to the enhancement of gene expression and immunogenicity of HPV 6b E7 gene.
Zhao
Weiming
W
Department of Microbiology, School of Medicine, Shandong University, Jinan 250012, China.
Yu
Xiuping
X
Zhou
Yabin
Y
Bian
Jifeng
J
Jia
Jihui
J
Luan
Yi
Y
Qi
Mei
M
Sun
Xinliu
X
Wang
Hong
H
D000818:Animals; D002460:Cell Line; D003062:Codon / Q000235:genetics*; D005260:Female; D015870:Gene Expression; D005814:Genes, Viral / Q000235:genetics*; D005822:Genetic Vectors; D051379:Mice; D027383:Papillomaviridae / Q000235:genetics*; D053918:Papillomavirus Vaccines; D014162:Transfection; D019444:Vaccines, DNA / Q000276:immunology*; D014764:Viral Proteins / Q000096:biosynthesis* / Q000235:genetics; D014765:Viral Vaccines / Q000276:immunology*
D003062:Codon; D053918:Papillomavirus Vaccines; D019444:Vaccines, DNA; D014764:Viral Proteins; D014765:Viral Vaccines