[Construction of adeno-associated virus vector containing human papilloma virus 16 E7 gene and its expression in eukaryotic cells].
OBJECTIVE
To construct recombinant adeno-associated virus vector containing human papilloma virus (HPV) 16E7 gene and identify its effectiveness of expression in eukaryotic cell.


METHODS
Recombinant adeno-associated virus particles containing HPV16E7 gene were generated by co-transfection of plasmids pAAV-MCS-E7, pAAV-RC and pAAV-helper into HEK293 cells. The viral particles were collected and purified. The vector titer was measured by southern blot. After the eukaryotic cells were transfected by virus particles, RT-PCR and western blot analysis were performed to identify rAAV expression.


RESULTS
The recombinant adeno-associated virus vector containing HPV16E7 gene was correctly constructed. The vector titer measured by southern blot was approximately 1 x 10(11)/ml. After the eukaryotic cells were transfected by the recombinant adeno-associated virus vector, the expression of HPV16E7 gene was identified by RT-PCR and western blot.


CONCLUSIONS
Recombinant adeno-associated virus HPV16E7 vector is successfully constructed and can stably express in eukaryotic cells.
Zhang
Juan
J
Department of Gynecology, Second Affiliated Hospital, Sun Yat-sen University, Guangzhou 510120, China.
Lin
Zhong-qiu
ZQ
Peng
Yong-pai
YP
Zhang
Bing-zhong
BZ
Zhou
Hui
H
Yang
Qing-yuan
QY
D000256:Adenoviridae / Q000235:genetics*; D002478:Cells, Cultured; D005260:Female; D005822:Genetic Vectors; D052162:Human papillomavirus 16 / Q000235:genetics*; D006801:Humans; D007668:Kidney / Q000166:cytology; D050725:Papillomavirus E7 Proteins / Q000235:genetics* / Q000378:metabolism
D050725:Papillomavirus E7 Proteins; C059731:oncogene protein E7, Human papillomavirus type 16