# SOMATIC DNA RECOMBINATION IN ANTIBODY GENES

> **NIH NIH R01** · UNIVERSITY OF CALIFORNIA BERKELEY · 1989 · $226,208

## Abstract

The purpose of this proposed research is to analyze the molecular
mechanisms of somatic DNA recombination in immunoglobulin (Ig)
and T-cell receptor (TCR) genes.  We will define each individual
step of variable (V) region gene rearrangement, and characterize
the enzymatic machinary for the recombination.

We have identified a pre-B cell nuclear protein which specifically
interacts with the recombination signal sequences (RSS's).  This
protein is assumed to be a DNA binding component of a putative
recombinase.  In the proposed research, we will purify the RSS
binding protein with various chromatography columns including a
DNA cellulose (affinity) column.  The peptide sequence will be
determine for the binding protein, and oligonucleotide probes will
be snythesized for the cDNA cloning.  A cDNA library from the
pre-B cell line, 38B9, will be made with the expression vector,
phage lambda gtll.  The library will be screened with the
oligonucleotide probes to isolate the gene coding for the RSS
binding protein.

We have identified the TG-endonuclease which introduces a
double-strand cut adjacent to the recombination signal,
CACTGTG.  This nuclease is found in the recombination-
competent pre-B cell line, 38B9, and appears to be a good
candidate for the cleavage activity of the recombinase.  We will
purify the TG-endonuclease and isolate the cDNA clone in order
to study the structure and expression of the nuclease.

We have demonstrated that two sets of recombination signal
sequences, CACTGTG and GGTTTTTGT, were sufficient to cause
the V-J type of recombination when they were introduced into the
recombination competent pre-B cells.  With various mutant RSS
substrates, we will determine the structural requirements of
substrate DNA for the V-J type of recombination.

We have characterized the extrachromosomal circular DNA which
is an excision product of the gene rearrangement.  We will extend
our study of the circular DNA to the developmental regulation of
the Ig and TCR gene rearrangement.

It is hoped that the proposed research will clarify not only the
basic process of DNA recombination, but the molecular
mechanisms of immuno-deficiencies and leukemogenesis caused
by DNA rearrangement.

## Key facts

- **NIH application ID:** 3128214
- **Project number:** 5R01AI018790-08
- **Recipient organization:** UNIVERSITY OF CALIFORNIA BERKELEY
- **SAM.gov UEI:** GS3YEVSS12N6
- **Principal Investigator:** HITOSHI SAKANO
- **Activity code:** R01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 1989
- **Award amount:** $226,208
- **Award type:** 5
- **Project period:** 1982-08-01 → 1993-08-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/3128214

## Citation

> US National Institutes of Health, RePORTER application 3128214, SOMATIC DNA RECOMBINATION IN ANTIBODY GENES (5R01AI018790-08). Retrieved via MORVS 2026-10-25 from https://api.morvs.ai/grant/nih/3128214. Licensed CC0.

---

*[NIH grants dataset](/datasets/nih-grants) · CC0 1.0*