A human t-PA mutant cDNA cassette knocked in the murine fgfr-4 locus targeting for mammary gland expression.
The expression of foreign gene in transgenic animals produced by pronuclear microinjection is often confounded by the position effects caused by not only the nature of chromosomal integration site but also the number and arrangement of multiple transgene copies. Gene targeting provides a new way to overcome these inhibitions by introducing single-copy transgene into a chosen site. The choice of a good chromosomal site will favor transgene expression in a predictable fashion. In this study, we tested a new site (fgfr-4) for foreign gene integration and expression. A t-PA mutant (t-PAm) expression cassette under bovine alphas1-casein regulatory sequences was efficiently knocked-in fgfr-4 site through homologous recombination. The t-PAm was expressed in the milk of all targeted mice. Our experiment indicates that the fgfr-4 may be a candidate site for knocking foreign gene to make transgenic animals.
Lin
Fu-Yu
FY
Beijing Institute of Biotechnology, No.20 Dongdajie Street, Fengtai District, Beijing 100071, China.
Chen
Hong-Xing
HX
Cheng
Xuan
X
Xiao
Yang
Y
Deng
Ji-Xian
JX
Huang
Pei-Tang
PT
D000818:Animals; D001483:Base Sequence; D017931:DNA Primers; D018076:DNA, Complementary / Q000235:genetics*; D005260:Female; D005838:Genotype; D006801:Humans; D008297:Male; D042361:Mammary Glands, Human / Q000378:metabolism*; D051379:Mice; D008810:Mice, Inbred C57BL; D008892:Milk; D016254:Mutagenesis, Insertional; D051499:Receptor, Fibroblast Growth Factor, Type 4; D017468:Receptors, Fibroblast Growth Factor
D017931:DNA Primers; D018076:DNA, Complementary; D017468:Receptors, Fibroblast Growth Factor; C496371:FGFR4 protein, human; C496372:Fgfr4 protein, mouse; D051499:Receptor, Fibroblast Growth Factor, Type 4