[Construction of recombinant adenovirus vector carrying secondary lymphoid organ chemokine].
OBJECTIVE
To construct a recombinant adenovirus vector carring the SLC gene for further studies on gene therapy for carcinoma.


METHODS
The SLC gene was amplified from the pORF5 vector with polymerase chain reaction (PCR) technique. The amplified gene was cloned into the pENTR11 vector. With the pENTR11-SLC plasmid and the backbone plasmid pAd/CMV/V5-DEST, the homologous recombination reaction took place in vitro. The reaction mixture was transferred into TOP10 E. coli strains (without F' episome). The recombination adenovirus plasmid was then generated. The recombinant adenoviruses were packaged and amplified in 293A cells.


RESULTS
The SLC gene was successfully cloned into the pAd/CMV/V5-DEST plasmid and the recombinant adenoviruses carrying SLC gene were detected by PCR, with a viral titer of 2. 6 X 10(8) pfu/mL.


CONCLUSIONS
The constructed recombinant adenovirus can introduce SLC gene into tumor tissues, which provides a foundation for the study of antitumor efficacy of SLC.
Chen
Ping
P
Key Laboratory of Bio-resources and Eco-environment, Ministry of Education, College of Life Science, Sichuan University, Chengdu 610064, China.
Yang
Ling-lin
LL
Hu
Huo-zhen
HZ
D000256:Adenoviridae / Q000235:genetics* / Q000502:physiology; D054421:Chemokine CCL21 / Q000235:genetics*; D004274:DNA, Recombinant / Q000235:genetics*; D005818:Genetic Engineering / Q000379:methods*; D005822:Genetic Vectors / Q000235:genetics*; D006367:HeLa Cells; D006801:Humans; D010957:Plasmids / Q000235:genetics; D016133:Polymerase Chain Reaction; D019562:Viral Load
D054421:Chemokine CCL21; D004274:DNA, Recombinant