Expression and stable germline transmission of neomycin-resistance gene in transgenic mice.
Transgenic mice were produced to study the expression of amino-3' glycosyl phosphotransferase gene (neomycin resistance gene) in the embryonic fibroblast cells. A 1.9 Kb linear fragment of neomycin resistance gene under the control of pPGK promoter was microinjected into the pronucleus of mouse embryos. Out of 64 potential founders born, 5 were identified to be transgenic by the polymerase chain reaction (PCR) and southern hybridization. Multiple mice from first and second generation from two transgenic founders (N-10 and N-32) were analysed to determine the germline transmission. It was found to be 24.6 and 71.4% in first and second generation respectively. Results were also further confirmed by RT-PCR, sequencing and in vitro bioassays.
Kaur
G
G
Embryo Biotechnology Laboratory, National Institute of Immunology, New Delhi, India.
Gulati
N
N
Mandal
A
A
Babu
D S
DS
Anand
R
R
Garg
L
L
Totey
S M
SM
D000818:Animals; D001483:Base Sequence; D017931:DNA Primers; D004352:Drug Resistance, Microbial / Q000235:genetics; D005260:Female; D005854:Germ Cells; D019868:Kanamycin Kinase / Q000235:genetics*; D051379:Mice; D008810:Mice, Inbred C57BL; D008808:Mice, Inbred CBA; D008822:Mice, Transgenic; D009355:Neomycin / Q000494:pharmacology
D017931:DNA Primers; D019868:Kanamycin Kinase; D009355:Neomycin