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

Chi Hua Chiu, Chris T. Amemiya, Janet L. Carr, Jaya Bhargava, Joseph K. Hwang, Cooduvalli S. Shashikant, Frank H. Ruddle, Günter P. Wagner

Research output: Contribution to journalArticlepeer-review

15 Scopus citations

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.

Original languageEnglish (US)
Pages (from-to)105-109
Number of pages5
JournalDevelopment Genes and Evolution
Volume210
Issue number2
DOIs
StatePublished - Jan 1 2000

Keywords

  • AbDb-like Hox genes
  • Cis-regulatory sequences
  • Homologous recombination
  • P1/PAC clones
  • Reporter gene

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