# TARGETS OF THE MOUSE HOMEODOMAIN PROTEIN, HOX-1.4

> **NIH NIH F32** · COLUMBIA UNIV NEW YORK MORNINGSIDE · 1993 · $28,600

## Abstract

The research proposed is designed to identify specific genomic DNA target
sequences which bind the mouse homeodomain protein Hox 1.4, a protein which
may function as a transcription factor.  The Hox 1.4 gene is expressed in
the mid-gestation embryo and in meiotic and post-meiotic male germ cells.
Our plan of attack includes a series of protein/DNA binding assays and a
genetic selection protocol to identify genomic targets of Hox 1.4 protein
binding.  The first approach will utilize genomic DNA fragments ligated to
polymerase chain reaction (PCR) "catch linkers".  The DNA will be incubated
with the Hox 1.4 protein (produced in a Baculovirus expression system) to
allow protein/DNA binding.  DNA/protein complexes will be isolated by
standard immunoselection protocols using anti-Hox 1.4 antibodies.  The
selected DNA is amplified by PCR, and protein binding and immunoselection
are repeated to ensure isolation of specific sequences.  The DNA of
interest will be cloned, sequenced, and identified.  In our second
approach, Hox 1.4 DNA binding sequences (the homeodomain) are fused in
frame with the transcription activator sequences of GAL4 and cloned into an
expression plasmid with a selectable marker.  Another plasmid has mouse
genomic sequences cloned next to a reporter gene with a weak promoter.  Co-
transfection of the plasmids into yeast should result in clones producing
the Hox 1.4/GAL4 fusion protein which will bind to the mouse genomic
sequences and activate expression of the reporter gene.  These sequences
will be used as probes to identify the genes that Hox 1.4 binds.  These
experiments will identify specific DNA sequences which bind the Hox 1.4
protein and will enable us to determine which downstream genes may be
regulated by Hox 1.4, providing information on molecular events and gene
regulation during spermatogenesis.

## Key facts

- **NIH application ID:** 2195923
- **Project number:** 5F32HD007605-02
- **Recipient organization:** COLUMBIA UNIV NEW YORK MORNINGSIDE
- **SAM.gov UEI:** F4N1QNPB95M4
- **Principal Investigator:** STUART E RAVNIK
- **Activity code:** F32 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 1993
- **Award amount:** $28,600
- **Award type:** 5
- **Project period:** 1993-06-01 → —

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/2195923

## Citation

> US National Institutes of Health, RePORTER application 2195923, TARGETS OF THE MOUSE HOMEODOMAIN PROTEIN, HOX-1.4 (5F32HD007605-02). Retrieved via MORVS 2026-10-25 from https://api.morvs.ai/grant/nih/2195923. Licensed CC0.

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