Your browser doesn't support javascript.
loading
Optimized expression, preparation of human papillomavirus 16 L2E7 fusion protein and its inhibitory effect on tumor growth in mice / 生物工程学报
Chinese Journal of Biotechnology ; (12): 566-576, 2015.
Article in Chinese | WPRIM | ID: wpr-240618
ABSTRACT
HPV16 L2E7 is a fusion protein used for therapeutical vaccine targeting HPV virus. To increase its expression in Escherichia coli, we optimized the codon usage of HPV16 l2e7 gene based on its codon usage bias. The optimized gene of HPV16 sl2e7 was cloned into three different vectors pGEX-5X-1, pQE30, ET41a, and expressed in JM109, JM109 (DE3) and BL21 (DE3) lines separately. A high expression line was selected with pET41a vector in BL21 (DE3) cells. After optimization of the growth condition, including inoculation amount, IPTG concentration, induction time and temperature, the expression level of HPV16 L2E7 was increased from less than 10% to about 28% of total protein. HPV16 L2E7 protein was then purified from 15 L culture by means of SP Sepharose Fast Flow, Q Sepharose Fast Flow and Superdex 200 pg. After renaturing, HPV16 L2E7 protein with ≥ 95% purity was achieved, which was confirmed via SDS-PAGE gel and Western blotting. The combined use of purified HPV16 L2E7 and CpG helper has shown clear inhibition of tumor growth in mice injected with tumor cells, with six out of eight mice shown no sign of tumor. This study lays a solid foundation for a new pipeline of large-scale vaccine production.
Subject(s)
Full text: Available Index: WPRIM (Western Pacific) Main subject: Recombinant Fusion Proteins / Codon / Oncogene Proteins, Viral / Capsid Proteins / Therapeutic Uses / Electrophoresis, Polyacrylamide Gel / Escherichia coli / Human papillomavirus 16 / Papillomavirus E7 Proteins / Papillomavirus Vaccines Limits: Animals Language: Chinese Journal: Chinese Journal of Biotechnology Year: 2015 Type: Article

Similar

MEDLINE

...
LILACS

LIS

Full text: Available Index: WPRIM (Western Pacific) Main subject: Recombinant Fusion Proteins / Codon / Oncogene Proteins, Viral / Capsid Proteins / Therapeutic Uses / Electrophoresis, Polyacrylamide Gel / Escherichia coli / Human papillomavirus 16 / Papillomavirus E7 Proteins / Papillomavirus Vaccines Limits: Animals Language: Chinese Journal: Chinese Journal of Biotechnology Year: 2015 Type: Article