Volume 3, Issue 1 (March 2001) 3, 49–53;
Introduction of DT40 cells into chick embryos
M. Toba, F. Ebara, H. Furuta, Y. Matsushimal, Y. Kitagawa, N. Fujihara
Graduate Program for Regulation of Biological Signals, Graduate School of Bioagricultural Science, Nagoya University, Nagoya, 464-8601 Japan
Advance online publication 1 March 2001
Abstract |
Aim: 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 -galactosidase: -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 -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. Conclusion: 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
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