Preliminary application of a mosquito densovirus-mediated artificial intron in vitro and in vive of mosquito / 病毒学报
Chinese Journal of Virology
;
(6): 379-384, 2010.
Article
in Chinese
| WPRIM
| ID: wpr-286108
ABSTRACT
An artificial intron consisting of the 5'-donor site (from the first intron of the human beta-globin gene) and the 3'-acceptor site (from the intron of an immunoglobulin gene heavy chain variable region) was obtained with a splice overlap extension PCR and was then inserted in frame into the coding sequence of nostructural protein NS1 gene fused to GFP gene in a recombinant mosquito densovirus plasmid p7NS1-GFP. The constructed plasmid was named as p7NS1-Intron-GFP. The plasmid p7NS1-Intron-GFP was co transfected with the helper plasmid pUCA into C6/36 cells, then the packaged recombinant and wild type viruses were purified and recovered. The second-instars of Aedes albopictus larvae were exposed to recombinant and wild type virus mixed stock. The high level GFP expression in C6/36 cells and larvae was observed under fluorescence microscope, indicating that the inserted artificial intron exerted its normal function in self-splicing both in vitro and in vivo. This study laid a foundation for application of an artificial intron in insect cells and development of new strategy for genetic engineering technology of mosqtuito and its pathogens.
Full text:
Available
Index:
WPRIM (Western Pacific)
Main subject:
Virology
/
Introns
/
Cell Line
/
Polymerase Chain Reaction
/
Densovirus
/
Green Fluorescent Proteins
/
Genetics
/
Metabolism
/
Culicidae
Limits:
Animals
Language:
Chinese
Journal:
Chinese Journal of Virology
Year:
2010
Type:
Article
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