caa a22 4500 47576837X CHVBK 20180406123612.0 cr unu---uuuuu 170329e20000101xx s 000 0 eng 10.1007/s004270050016 doi (NATIONALLICENCE)springer-10.1007/s004270050016 A recombinogenic targeting method to modify large-inserts for cis -regulatory analysis in transgenic mice: construction and expression of a 100-kb, zebrafish Hoxa-11b-lacZ reporter gene [Elektronische Daten] [C-H. Chiu, C. T. Amemiya, J. L. Carr, J. Bhargava, J. K. Hwang, C. S. Shashikant, F. H. Ruddle, G. P. Wagner] Abstract : The identification of cis-sequences responsible for spatiotemporal patterns of gene expression often requires the functional analysis of large genomic regions. In this study a 100-kb zebrafish Hoxa-11b-lacZ reporter gene was constructed and expressed in transgenic mice. PAC clone 10-O19, containing a portion of the zebrafish HoxA-b cluster, was captured into the yeast-bacterial shuttle vector, pPAC-ResQ, by recombinogenic targeting. A lacZ reporter gene was then inserted in-frame into exon 1 of the zfHoxa-11b locus by a second round of recombinogenic targeting. Expression of the zfHoxa-11b-lacZ reporter gene in 10.5 d.p.f. transgenic mouse embryos was observed only in the posterior portion of the A-P axis, in the paraxial mesoderm, neural tube, and somites. These findings demonstrate the utility of recombinogenic targeting for the modification and expression of large inserts captured from P1/PAC clones. Springer-Verlag Berlin Heidelberg, 2000 Key words Cis-regulatory sequences nationallicence P1/PAC clones nationallicence Homologous recombination nationallicence Reporter gene nationallicence AbDb-like Hox genes nationallicence Chiu C-H Department of Ecology and Evolutionary Biology, Osborn Memorial Laboratory 327, Yale University, P.O. Box 208106, New Haven, CT 06520-8106, USA e-mail: chi-hua.chiu@yale.edu Tel.: +1-203-4329999, Fax: +1-203-4323870, US aut Amemiya C. T. Center for Human Genetics, Boston University School of Medicine, 700 Albany Street, W408, Boston, MA 02118, USA, US aut Carr J. L. Genaissance Pharmaceuticals Inc., 5 Science Park, New Haven, CT 06511, USA, US aut Bhargava J. Genaissance Pharmaceuticals Inc., 5 Science Park, New Haven, CT 06511, USA, US aut Hwang J. K. Department of Obstetrics and Gynecology, Yale University, School of Medicine, P.O. Box 208063, New Haven, CT 06520, USA, US aut Shashikant C. S. Department of Dairy and Animal Science, College of Agricultural Sciences, Pennsylvania State University, 324 Henning Building, University Park, PA 16802-3503, USA, US aut Ruddle F. H. Departments of Molecular, Cellular, and Developmental Biology, Kline Biology Tower, Yale University, P.O. Box 208103, New Haven, CT 06520, USA, US aut Wagner G. P. Department of Ecology and Evolutionary Biology, Osborn Memorial Laboratory 327, Yale University, P.O. Box 208106, New Haven, CT 06520-8106, USA e-mail: chi-hua.chiu@yale.edu Tel.: +1-203-4329999, Fax: +1-203-4323870, US aut https://doi.org/10.1007/s004270050016 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/s004270050016 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Chiu C-H Department of Ecology and Evolutionary Biology, Osborn Memorial Laboratory 327, Yale University, P.O. Box 208106, New Haven, CT 06520-8106, USA e-mail: chi-hua.chiu@yale.edu Tel.: +1-203-4329999, Fax: +1-203-4323870, US aut NATIONALLICENCE 700 1- Amemiya C. T. Center for Human Genetics, Boston University School of Medicine, 700 Albany Street, W408, Boston, MA 02118, USA, US aut NATIONALLICENCE 700 1- Carr J. L. Genaissance Pharmaceuticals Inc., 5 Science Park, New Haven, CT 06511, USA, US aut NATIONALLICENCE 700 1- Bhargava J. Genaissance Pharmaceuticals Inc., 5 Science Park, New Haven, CT 06511, USA, US aut NATIONALLICENCE 700 1- Hwang J. K. Department of Obstetrics and Gynecology, Yale University, School of Medicine, P.O. Box 208063, New Haven, CT 06520, USA, US aut NATIONALLICENCE 700 1- Shashikant C. S. Department of Dairy and Animal Science, College of Agricultural Sciences, Pennsylvania State University, 324 Henning Building, University Park, PA 16802-3503, USA, US aut NATIONALLICENCE 700 1- Ruddle F. H. Departments of Molecular, Cellular, and Developmental Biology, Kline Biology Tower, Yale University, P.O. Box 208103, New Haven, CT 06520, USA, US aut NATIONALLICENCE 700 1- Wagner G. P. Department of Ecology and Evolutionary Biology, Osborn Memorial Laboratory 327, Yale University, P.O. Box 208106, New Haven, CT 06520-8106, USA e-mail: chi-hua.chiu@yale.edu Tel.: +1-203-4329999, Fax: +1-203-4323870, US aut Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer