The 5'-flanking region of the murine epididymal protein of 17 kilodaltons gene targets transgene expression in the epididymis.
A murine epididymal retinoic-acid-binding protein (mE-RABP) is specifically expressed in the mid/distal caput epididymidis and is androgen regulated. The murine epididymal protein of 17 kDa (mEP17) gene, a novel gene homologous to mE-RABP, is located within 5 kb of the 5'-flanking region of the mE-RABP gene. In contrast, expression of the mEP17 gene is restricted to the initial segment and regulated by factor(s) contained in testicular fluid. To identify cis-DNA regulatory element(s) involved in the tissue- and region-specific expression of the mEP17 gene in transgenic mice, we have studied the expression of a transgene containing 5.3 kb of the 5'-flanking region of the mEP17 gene (5.3mEP17) linked to chloramphenicol acetyltransferase (CAT) reporter gene. Significant caput epididymidis-specific CAT activity was detected in transgenic mouse lines; and CAT gene expression is restricted to the initial segment, as is the expression of the endogenous mEP17 gene. Ontogenic expression and testicular factor dependency also mimic that of endogenous mEP17 gene. These results suggest that the 5.3mEP17 fragment contains all the information required for spatial and temporal expression in the mouse epididymis. The 5.3mEP17 fragment will be useful to express a foreign gene of interest in the epididymis in an initial segment-specific manner.
Suzuki
Kichiya
K
Center for Reproductive Biology Research and Department of Obstetrics and Gynecology, Vanderbilt University, School of Medicine, Nashville, Tennessee 37232-2633, USA. kichiya.suzuki@vanderbilt.edu
Araki
Yoshihiko
Y
Zhu
Mei-Ying
MY
Lareyre
Jean-Jacques
JJ
Matusik
Robert J
RJ
Orgebin-Crist
Marie-Claire
MC
D000818:Animals; D002352:Carrier Proteins / Q000235:genetics*; D015500:Chloramphenicol O-Acetyltransferase / Q000235:genetics; D004822:Epididymis / Q000737:chemistry* / Q000378:metabolism*; D015870:Gene Expression; D017930:Genes, Reporter; D017403:In Situ Hybridization; D054834:Lipocalins; D008297:Male; D051379:Mice; D008822:Mice, Transgenic; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000235:genetics; D012333:RNA, Messenger / Q000032:analysis; D018168:Receptors, Retinoic Acid / Q000235:genetics; D011993:Recombinant Fusion Proteins; D012045:Regulatory Sequences, Nucleic Acid; D015183:Restriction Mapping; D019076:Transgenes / Q000235:genetics*
D002352:Carrier Proteins; C423382:Lcn8 protein, mouse; D054834:Lipocalins; D010446:Peptide Fragments; D012333:RNA, Messenger; D018168:Receptors, Retinoic Acid; D011993:Recombinant Fusion Proteins; D015500:Chloramphenicol O-Acetyltransferase