Format

Send to

Choose Destination
Zhonghua Yu Fang Yi Xue Za Zhi. 2010 Aug;44(8):721-5.

[Characterization and secreted expression of dengue virus type I-IV envelope glycoprotein domain III in Pichia pastoris].

[Article in Chinese]

Author information

1
Clinical Laboratory, Zhujiang Hospital, Southern Medical University, Guangzhou 510282, China.

Abstract

OBJECTIVE:

To achieve secretory and extracellular production of recombinant dengue virus serotypes I-IV envelope glycoprotein domain III (DENV-1-4 EDIII) in Pichia pastoris.

METHODS:

EDIII genes of DENVI-IV were amplified and cloned into vector pPIC9K, respectively. These recombinant plasmids were then linearized and transferred into Pichia pastoris strain GS115. Clones highly produced in 4.0 mg/ml G418 were amplified and induced by methanol to achieve the secreted recombinant proteins. Ni-NTA agarose beads were used for purification, while SDS-PAGE and Western blotting were used for identification.

RESULTS:

The recombinant plasmids pPIC9K-DENV-1-4 EDIII were constructed and successfully transferred into Pichia pastoris strain GS115. The recombinant EDIII proteins were expressed in a secretory way with the molecular weight about 12 × 10(3) and specifically identified by anti-His monoclonal antibody and anti-DENVI-IV mice sera.

CONCLUSION:

DENVI-IV EDIII proteins are successfully achieved from Pichia pastoris expression system and could be used for development of dengue vaccines, diagnostic reagents and study of biological function of the E protein.

PMID:
21055023
[Indexed for MEDLINE]

Supplemental Content

Full text links

Icon for Chinese Medical Association Publishing House Ltd.
Loading ...
Support Center