[Development of plasmid-based transgenic mice carrying with target gene xylE].
OBJECTIVE
To develop a transgenic mouse model carrying with plasmid pUC118NX integrated into its genomic DNA for detecting mutagenesis.


METHODS
DNA of plasmid UC118NX in target gene xy1E was injected microscopically into male protonucleus of 376 mouse spermatova, and 225 survival spermatova were transferred into the oviducts in both sides of 11 pseudopregnant female mice to develop their offspring. The genomic DNA in survival young mice were analyzed with polymerase chain reaction (PCR)-Southern blot, plasmid transformation test and endonuclease-digestion.


RESULTS
Seven pseudopregnant mice got pregnant, and 29 offspring were delivered and 25 survived of which 18 were identified with positive for PCR-Southern blot (72%). The two stout male mice with intact integration of plasmid pUC118NX in their genome were finally chosen as founders to detect gene mutation in vivo and establish transgenic mouse lineages.


CONCLUSIONS
Transgenic C57BL/6J mice integrated with plasmid pUC118NX into their genomic DNA have been successfully developed.
Cao
J
J
Department of Hygienic Toxicology The Second Military Medical University, Shanghai 200433.
Chen
J
J
Yin
M
M
Cheng
G
G
Huang
J
J
Mao
Y
Y
Xu
S
S
D000818:Animals; D015139:Blotting, Southern; D050557:Catechol 2,3-Dioxygenase; D049308:Dioxygenases; D051379:Mice; D008810:Mice, Inbred C57BL; D008822:Mice, Transgenic; D008845:Microinjections; D009152:Mutagenicity Tests; D010105:Oxygenases / Q000235:genetics*; D010957:Plasmids; D016133:Polymerase Chain Reaction
D010105:Oxygenases; D049308:Dioxygenases; D050557:Catechol 2,3-Dioxygenase