[Preliminary application of a mosquito densovirus-mediated artificial intron in vitro and in vive of mosquito].
Bing Du Xue Bao
; 26(5): 379-84, 2010 Sep.
Article
in Zh
| MEDLINE
| ID: mdl-21043138
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.
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Collection:
01-internacional
Database:
MEDLINE
Main subject:
Introns
/
Densovirus
/
Culicidae
Limits:
Animals
Language:
Zh
Journal:
Bing Du Xue Bao
Journal subject:
VIROLOGIA
Year:
2010
Document type:
Article
Affiliation country:
China