Distribution and expression of recombinant plasmid encoding chicken interleukin-2.
A plasmid DNA that encodes chicken interleukin-2 (pCI-ChIL-2-EGFP) was investigated for its distribution and expression after intramuscular (i.m.) injection in chickens. After the i.m. injection, serum distribution was detectable from 2 h post inoculation (p.i.), peaked at 8 h p.i., and disappeared at 7 days p.i. The plasmid DNA was also observed in several organs including heart, liver, lung, spleen, bursa and inoculated muscle at different time points, but at 19 days p.i. the plasmid DNA was not found in any organ except inoculated muscle. Fluorescence of enhanced green fluorescent protein (EGFP) was found in cytoplasm and nucleus of cultured Vero cells, chicken embryo fibroblasts and peripheral blood lymphocytes, which were transfected in vitro with the plasmid DNA or in vivo with Lipofectamine. The expression profile of the fusion gene (ChIL-2-EGFP) in vivo was measured by RT-PCR, ELISA and fluorescence microscopy. The EGFP expression was detected from 8 h p.i. to 14 days p.i. and peaked at 5 days p.i., when the number of EGFP-expression myocytes was about 5% in the injected site. These results demonstrate that intramuscular administration of plasmid DNA leads to widespread distribution and long-term expression in vivo.
You
Z Q
ZQ
Institute of Preventive Veterinary Medicine, Zhejiang University, Hangzhou, China.
Yu
L
L
Zhang
C Z
CZ
Li
L
L
Lu
M J
MJ
Mao
Z J
ZJ
Liu
Y
Y
Chu
W Y
WY
D000818:Animals; D002642:Chick Embryo; D002645:Chickens / Q000276:immunology / Q000378:metabolism*; D002522:Chlorocebus aethiops; D003001:Cloning, Molecular; D004274:DNA, Recombinant / Q000008:administration & dosage* / Q000235:genetics / Q000276:immunology / Q000493:pharmacokinetics; D004797:Enzyme-Linked Immunosorbent Assay / Q000662:veterinary; D007376:Interleukin-2 / Q000096:biosynthesis / Q000097:blood / Q000235:genetics* / Q000276:immunology; D008856:Microscopy, Fluorescence / Q000662:veterinary; D010957:Plasmids / Q000235:genetics / Q000378:metabolism / Q000493:pharmacokinetics*; D011897:Random Allocation; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000662:veterinary; D013047:Specific Pathogen-Free Organisms; D014162:Transfection / Q000662:veterinary; D014709:Vero Cells
D004274:DNA, Recombinant; D007376:Interleukin-2; D011993:Recombinant Fusion Proteins