Aim:To examine the transfection of exogenous genes into chick embryos,applying the characteristics of avianleukosis vires(ALV)-induced chicken B cell line DT40 to the production of chimeric birds.Methods:The DT40cells...Aim:To examine the transfection of exogenous genes into chick embryos,applying the characteristics of avianleukosis vires(ALV)-induced chicken B cell line DT40 to the production of chimeric birds.Methods:The DT40cells incorporated with exogenous gene(lacZ constructs encoding Escherichia coliβ-galactosidase:β-gal)were intro-duced into chick embryos by the injection of cells into stage X blastoderm.Manipulated eggs were incubated for 3(trial1)or 6(trial 2)days,and the expression of lacZ DNA was detected by a histochemical staining method ofβ-galactosi-dase and polymerase chain reaction(PCR)analysis.Results:The survival rates of the manipulated embryos incu-bated 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,forthe survived embryos.Conclusion:The rate of embryonic viability and expression rate of introduced genes were notso high,but it suggested the possibility of utilizing the DT40 cells as a vector for carrying exogenous genes into chickembryos.展开更多
基金financially supported by the Ministry of Education,Science and Culture,Japanthe Society for the Promotion of Science(JSPS)+1 种基金the Sumitomo Foundationthe Nissan Science Foundation
文摘Aim:To examine the transfection of exogenous genes into chick embryos,applying the characteristics of avianleukosis vires(ALV)-induced chicken B cell line DT40 to the production of chimeric birds.Methods:The DT40cells incorporated with exogenous gene(lacZ constructs encoding Escherichia coliβ-galactosidase:β-gal)were intro-duced into chick embryos by the injection of cells into stage X blastoderm.Manipulated eggs were incubated for 3(trial1)or 6(trial 2)days,and the expression of lacZ DNA was detected by a histochemical staining method ofβ-galactosi-dase and polymerase chain reaction(PCR)analysis.Results:The survival rates of the manipulated embryos incu-bated 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,forthe survived embryos.Conclusion:The rate of embryonic viability and expression rate of introduced genes were notso high,but it suggested the possibility of utilizing the DT40 cells as a vector for carrying exogenous genes into chickembryos.
