Construction and identification of recombinant adenovirus vector containing human TLR4 extracellular domain gene
LIU Jing
LI Qiang
SUN Shouxun
LI Ling
CHAI Ruonan
PU Xiaoyun
Abstract：Objective To construct and identify human TLR4 extracellular recombinant adenovirns. Methods TLR4 extra-cellular domain gent was amplified by PCR with pUCm-TLR4 plasmid as template, and then connected to the shuttle vector pAdTrack-CMV for constructing recombinant adenovirus shuttle plasmid pAdTrack-CMV- TLR4 by enzyme digestion. After sequencing, the plasma was linearized with pmeⅠ and cotransformed into E. coll. BJ5183 cells containing pAdEasy-I plasmid to undergo homologous recombina-tion. After been screened by selected media, the extracted plasmid of positive bacteria was transfected into 293 cells with liposome, and then identified by green fluorescence staining and PCR. Results The TLR4 extraccllular domain gene was inserted successfully into the recombinant adenovims vector, with titers of about 3.2×109 pfu/mL. Conclusion The recombinant adcnovirus vector of human TLR4 extracellular domain gene is successfully constructed, which would be a foundation of next research.
Keywords：TLR4- adenovirus vectorextracellular domain
Publication Date：2009-01-01
Online Publishing Date：2025-08-15(First online date of this platform, not the publication date of the document)
Pages：4( 457-460 )