[Construction of eukaryotic expressing plasmids for HA and HA1 of influenza A virus and their transient expression in HEK293 cells].
OBJECTIVE
To construct influenza A virus (A/PR/8/34) HA and HA1 eukaryotic expressing plasmids and study their expression in HEK293 cells.


METHODS
HA and HA1 genes were cloned by RT-PCR and then inserted into pcDNA3. 1 (+). After identification of restriction enzyme digestion, PCR and sequencing analysis, HA and HA1 eukaryotic expressing plasmids were transfected into HEK293 cells with PolyFect Transfection Reagent. Immunofluorescence assay was used to observe the transient expressing result.


RESULTS
It was confirmed that the construction of HA and HA1 eukaryotic expressing plasmids was made successfully. The stronger fluorescence signals were detected in transfected HEK293 cells with these two kinds of plasmids by immunofluorescence assay.


CONCLUSIONS
The experiment is a success in the construction of eukaryotic expressing plasmids for HA and HA1, thus providing a basis for further probing into the mechanism of virus infection and exploring DNA vaccine.
Zhang
Wei-Dong
WD
Medicine, Sichuan University, Chengdu 610041, China.
Ca
Kang
K
Li
Ming-Yuan
MY
Shi
Qiao-Fa
QF
Wang
Bao-Ning
BN
Jiang
Zhong-Hua
ZH
Li
Hong
H
D002460:Cell Line; D003001:Cloning, Molecular; D005057:Eukaryotic Cells / Q000378:metabolism*; D005822:Genetic Vectors; D019267:Hemagglutinin Glycoproteins, Influenza Virus / Q000096:biosynthesis* / Q000235:genetics; D006801:Humans; D009980:Influenza A virus / Q000235:genetics* / Q000378:metabolism; D007668:Kidney / Q000166:cytology / Q000378:metabolism*; D010957:Plasmids / Q000378:metabolism; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics; D014162:Transfection
D019267:Hemagglutinin Glycoproteins, Influenza Virus; D011994:Recombinant Proteins; C483468:hemagglutinin, human influenza A virus