# GENE REGULATION DURING EARLY EMBRYONIC DEVELOPMENT

> **NIH NIH R01** · BAYLOR COLLEGE OF MEDICINE · 1994 · $199,503

## Abstract

The broad objectives of this proposal are the identification,
characterization and functional analysis of embryonic regulatory proteins
and their corresponding genes that control early gene activity and thus,
elucidate the mechanism responsible for the establishment of the early
embryonic cell lineages during the cleavage stages in the sea urchin
embryo. The short term goals of the proposed research studies are the
purification of two embryonic regulatory factors (CF1 and EFl) that are
differentially distributed in time and space within the developing embryo,
the cloning and characterization of their corresponding genes, and the
functional significance of these regulators in ontogeny of the sea urchin
and other embryos. CFI is a maternal, in origin, nuclear protein that is
found exclusively in the ectodermal cells of the embryo. EF1 is an early
embryonic but not maternal nuclear protein that exhibits two distinct site
specific binding forms before and after gastrulation; the early one is
present in the ectoderm, whereas the late form is contained in the
meso-endodermal germ layer. Both factors have their target sites in the
regulatory region of the early cleavage activated cytoskeletal actin gene,
CyIIIb. Purification of the two proteins will be carried out by DNA
affinity chromatography using early and late embryonic nuclear extracts.
The amino acid sequence of the two proteins will be determined and used to
synthesize nucleotide probes which will allow the cloning of their
corresponding cDNAs. The structure of these proteins will be determined by
sequencing full length CDNA clones. The developmental pattern of
expression of the corresponding genes will be analyzed in time and space
by RNA blot and in situ hybridizations to embryos from different stages.
The protein localization within the embryo will be analyzed with the use
of antibodies raised against the embryonic or the recombinant proteins. We
will study the ontogenic significance of these regulatory factors and
their functional domains in vivo by introducing intact and mutant proteins
and test fusion genes, by microinjection, into developing sea urchin
embryos and Xenopus oocytes.

## Key facts

- **NIH application ID:** 2198433
- **Project number:** 5R01HD022055-07
- **Recipient organization:** BAYLOR COLLEGE OF MEDICINE
- **SAM.gov UEI:** FXKMA43NTV21
- **Principal Investigator:** CONSTANTIN N FLYTZANIS
- **Activity code:** R01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 1994
- **Award amount:** $199,503
- **Award type:** 5
- **Project period:** 1986-07-01 → 1995-06-30

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/2198433

## Citation

> US National Institutes of Health, RePORTER application 2198433, GENE REGULATION DURING EARLY EMBRYONIC DEVELOPMENT (5R01HD022055-07). Retrieved via MORVS 2026-10-25 from https://api.morvs.ai/grant/nih/2198433. Licensed CC0.

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