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ISSN 0582-9879 Acta Biochimica et Biophysica Sinica 2004, 36(7): 450–456 CN 31-1300/Q
A Human t-PA Mutant cDNA Cassette Knocked in the Murine fgfr-4 Locus Targeting for Mammary Gland Expression
Fu-Yu LIN#,
Hong-Xing CHEN#, Xuan CHENG, Xiao YANG*, Ji-Xian DENG*, and Pei-Tang
HUANG*
Beijing Institute of Biotechnology, No.20 Dongdajie Street, Fengtai District, Beijing 100071, China
Abstract 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 as1-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.
Key words gene targeting;
fgfr-4; transgenic; mammary gland bioreactor; integration site; t-PA mutant
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Received: February 19, 2004 Accepted: May 17, 2004
This work was supported by a grant from the National High Technology Research and Development Program of China (2001AA213031)
#These two authors contributed equally to this work
*Corresponding authors:
Xiao YANG: Tel, 86-10-63895937; Fax, 86-10-63833521; E-mail, [email protected]
Ji-Xian DENG: Tel, 86-10-63858604; Fax, 86-10-63833521; E-mail, [email protected]
Pei-Tang HUANG: Tel, 86-10-66931007; Fax, 86-10-63833521; E-mail,
[email protected]