Introduction of DT40 cells into chick embryos
- Author:
Toba MARIKO
1
;
Ebara FUMIO
;
Furuta HIROKI
;
Matsushimal YUICHI
;
Kitagawa YASUO
;
Fujihara NOBORU
Author Information
1. Graduate School Kyushu University
- Keywords:
DT40 cells;
chick embryo;
lacZ;
polymerase chain reaction
- From:
Asian Journal of Andrology
2001;3(1):49-53
- CountryChina
- Language:Chinese
-
Abstract:
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 DT40cells incorporated with exogenous gene (lacZ constructs encoding Escherichia coli β-galactosidase: β-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 β-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. Conclusio: 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.
