Introduction of DT40 cells into chick embryos.
OBJECTIVE
To examine the transfection of exogenous genes into chick embryos, applying the characteristics of avian leukosis virus (ALV)-induced chicken B cell line DT40 to the production of chimeric birds.


METHODS
The DT40 cells incorporated with exogenous gene ( lacZ constructs encoding Escherichia coli beta-galactosidase: beta-gal) were introduced into chick embryos by the injection of cells into stage X blastoderm. Manipulated eggs were incubated for 3 (trial 1) or 6 (trial 2) days, and the expression of lacZ DNA was detected by a histochemical staining method of beta-galactosidase and polymerase chain reaction (PCR) analysis.


RESULTS
The survival rates of the manipulated embryos incubated for 3 days (stage 18-20: trial 1) and 6 days (stage 28, 30: trial 2) were about 42% and 38%, respectively. The expression rates of the lacZ gene in the embryos in the trials 1 and 2 were about 60% and 23%, respectively, for the survived embryos.


CONCLUSIONS
The rate of embryonic viability and expression rate of introduced genes were not so high, but it suggested the possibility of utilizing the DT40 cells as a vector for carrying exogenous genes into chick embryos.
Division of Bioresource and Bioenvironmental Sciences, Graduate School Kyushu University, Fukuoka, Japan.
M
M
Toba
F
F
Ebara
H
H
Furuta
Y
Y
Matsushimal
Y
Y
Kitagawa
N
N
Fujihara
D000818:Animals; D002460:Cell Line; D002478:Cells, Cultured; D002642:Chick Embryo / Q000201:enzymology*; D002678:Chimera / Q000235:genetics; D004247:DNA / Q000032:analysis* / Q000235:genetics; D004587:Electrophoresis, Agar Gel; D015870:Gene Expression; D016133:Polymerase Chain Reaction; D014162:Transfection; D001616:beta-Galactosidase / Q000235:genetics* / Q000378:metabolism
D004247:DNA; D001616:beta-Galactosidase