Construction and Expression of RNase-Resisting His-Tagged Virus-Like Particles Containing FluA/B mRNA / 病毒学报
Chinese Journal of Virology
;
(6): 629-633, 2015.
Article
in Chinese
| WPRIM
| ID: wpr-296237
ABSTRACT
To prepare virus-like particles containing FluA/B mRNA as RNA standard and control in Influenza RNA detection, the genes coding the coat protein and maturase of E. coli bacteriophage MS2 were amplified and cloned into D-pET32a vector. Then we inserted 6 histidines to MS2 coat protein by QuikChange Site-Directed Mutagenesis Kit to construct the universal expressing vector D-pET32a-CP-His. In addition, the partial gene fragments of FluA and FluB were cloned to the down-stream of expressing vector. The recombinant plasmid D-pET32a-CP-His-FluA/B was transformed to BL21 with induction by IPTG. The virus-like particles were purified by Ni+ chromatography. The virus-like particles can be detected by RT-PCR, but not PCR. They can be conserved stably for at least 3 months at both 4 degrees C and -20 degrees C. His-tagged virus-like particles are more stable and easier to purification. It can be used as RNA standard and control in Influenza virus RNA detection.
Full text:
Available
Index:
WPRIM (Western Pacific)
Main subject:
Influenza A virus
/
Influenza B virus
/
Ribonucleases
/
Virion
/
Recombinant Fusion Proteins
/
RNA, Messenger
/
RNA, Viral
/
Chemistry
/
Escherichia coli
/
Genetics
Language:
Chinese
Journal:
Chinese Journal of Virology
Year:
2015
Type:
Article
Similar
MEDLINE
...
LILACS
LIS