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Updated: Aug 9, 2026

Functional Cloning Using a Xenopus Oocyte Expression System
Published on: January 30, 2016
Cloning of a lymphoid-specific cDNA encoding a protein binding the regulatory octamer DNA motif
L M Staudt1, R G Clerc, H Singh
1Whitehead Institute for Biomedical Research, Cambridge, MA 02142.
Abstract:
An octamer DNA sequence plays a critical role in directing transcription of immunoglobulin genes in B lymphocytes. A new technique of direct binding of radioactive DNA was used to screen a complementary DNA expression library from the BJAB cell line in lambda gt11 phage to derive molecular cDNA clones representing a putative B lymphocyte-specific octamer binding protein. The plaques were screened with DNA containing four copies of the octamer sequence and positive phage recombinants were identified. The fusion protein produced on inducing a lysogen of one phage bound to a monomeric octamer probe. The cDNA insert from this phage hybridized to messenger RNA found in B lymphocytes, but not in most other cells. Thus, this cDNA derives from a gene (oct-2) that specifies an octamer binding protein expressed preferentially in B lymphocytes, proving that, for at least one gene, a cell-specific transcription factor exists and its amount is controlled through messenger RNA availability.
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