[Optimized expression of the L1 protein of human papillomavirus in Hansenula polymorpha].
The heterologously expressed L1 protein of human papilomavirus 16 can assembly into virus-like particles (VLPs), which has been used as prophylactic vaccine for cervical carcinoma. To express L1 protein in Hansenula polymorpha, we analyzed the codon usage of the native gene of L1 protein and redesigned the encoding sequence according to the codon bias of H. polymorpha. We used assembly PCR to synthesize the native gene HPV16L1-N and the codon optimized gene HPV16L1. The synthesized genes were cloned into pMOXZa-A vector to generate plasmids pMOXZ-HPV16N and pMOXZ-HPV16. The expression cassettes MOXp-HPV16L1(N)-AOXTT were cloned into YEp352 vector and transferred into H. polymorpha. After methanol inducement, the expression of L1 protein in H. polymorpha was detected from the codon optimized gene HPV16L1 rather than the native gene HPVI6L1-N. The parameters for induced cultivation for strain HP-U-16L with HPV16L1 were investigated in shaking flask cultures. After induced cultivation in YPM (pH 7.0) medium supplemented with methanol to a final concentration of 1.0% every 12 h at 37 degrees C for 72 h, the recombinant produced 78.6 mg/L of L1 protein. This work offers the possibility for the production of prophylactic vaccine for cervical carcinoma by H. polymorpha.
Li
Weiwei
W
Laboratory of Yeast Molecular Genetics and Breeding, Institute of Microbiology, Chinese Academy of Sciences, Beijing 100101, China.
He
Xiuping
X
Guo
Xuena
X
Zhang
Zhenying
Z
Zhang
Borun
B
D036022:Capsid Proteins / Q000096:biosynthesis* / Q000235:genetics; D003001:Cloning, Molecular; D003062:Codon / Q000235:genetics; D005822:Genetic Vectors / Q000235:genetics; D052162:Human papillomavirus 16 / Q000235:genetics*; D009856:Oncogene Proteins, Viral / Q000096:biosynthesis* / Q000235:genetics; D010843:Pichia / Q000235:genetics / Q000378:metabolism*; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics
D036022:Capsid Proteins; D003062:Codon; D009856:Oncogene Proteins, Viral; D011994:Recombinant Proteins; C063317:L1 protein, Human papillomavirus type 16