Prokaryotic expression of HPV 16L1 and optimization of expression conditions / 浙江大学学报·医学版
Journal of Zhejiang University. Medical sciences
;
(6): 395-398, 2010.
Artículo
en Chino
| WPRIM
| ID: wpr-319888
ABSTRACT
<p><b>OBJECTIVE</b>To construct the HPV16 L1 prokaryotic expression plasmid and to optimize its expression.</p><p><b>METHODS</b>A pair of primers was designed according to plasmid sequences of pGEX-KG and the HPV16L1 genes published by GeneBank. The DNA fragment of 1500 bp was amplified by PCR from the HPV recombinant plasmid with HPV16L1 gene, then cloned into pGEX-KG and transformed into the host E.coli strain JM109. The pGEX-KG-HPV16L1 plasmid was taken and transformed into BL21(DE3) for expression. Induced by IPTG at 37 degree, the expression product of HPV16L1 gene was identified by SDS-PAGE and Western blot.</p><p><b>RESULTS</b>HPV16L1 fusion protein was expressed successfully in the form of inclusion bodies. The molecular weight was 83 kD. Meanwhile, the optimum condition of HPV16L1 fusion protein expression was induced with 1.0 mmol*L(-1) IPTG for 4 h. The fusion protein reacted specifically with antibodies against HPV16L1.</p><p><b>CONCLUSION</b>The prokaryotic expression vector of HPV16L1 gene has been constructed and expressed in E.coli successfully.</p>
Texto completo:
Disponible
Índice:
WPRIM (Pacífico Occidental)
Asunto principal:
Proteínas Recombinantes de Fusión
/
Proteínas Oncogénicas Virales
/
Clonación Molecular
/
Vacunas contra el Cáncer
/
Proteínas de la Cápside
/
Escherichia coli
/
Papillomavirus Humano 16
/
Vectores Genéticos
/
Genética
/
Metabolismo
Idioma:
Chino
Revista:
Journal of Zhejiang University. Medical sciences
Año:
2010
Tipo del documento:
Artículo
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