[Cloning and expression of PAL gene of Legionella pneumophila].
OBJECTIVE
To develop DNA vaccine for Legionella pneumophila.


METHODS
PAL gene of Legionella pneumophila was amplified with PCR. The amplified DNA was ligated to pcDNA3.1(+) vector. The recombinant plasmid, which was identified by restriction analysis, PCR and sequence analysis, was named pcDNA3.1-PAL. The NIH3T3 cells were transfected with the recombinant plasmid pcDNA3.1-PAL by Lipofection. Transient and stable expression products of the PAL gene were detected by immunofluorescence and RT-PCR.


RESULTS
The recombinant plasmid pcDNA3.1-PAL expressed PAL protein in the eukaryotic cell NIH3T3.


CONCLUSIONS
This study has built a foundation for the development of PAL gene DNA vaccine for Legionella pneumophila.
Tian
Yu
Y
Department of Parasitology, West China School of Preclinical and Forensic Medicine, Sichuan University, Chengdu 610041, China.
Chen
Jian-ping
JP
Yang
Chun-lei
CL
Liu
Ming-jie
MJ
D000818:Animals; D003001:Cloning, Molecular; D015870:Gene Expression; D005818:Genetic Engineering; D007150:Immunohistochemistry; D016952:Legionella pneumophila / Q000235:genetics* / Q000276:immunology; D008074:Lipoproteins / Q000235:genetics* / Q000302:isolation & purification; D051379:Mice; D041681:NIH 3T3 Cells; D010957:Plasmids / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D019444:Vaccines, DNA / Q000235:genetics / Q000276:immunology
D008074:Lipoproteins; D019444:Vaccines, DNA