HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS		QUICK SEARCH:   [advanced] Author: Keyword(s): Year:  Vol:  Page:

J. Biol. Chem., Vol. 276, Issue 23, 19913-19920, June 8, 2001

This Article Full Text Full Text (PDF) All Versions of this Article: 276/23/19913    most recent M100448200v1 Alert me when this article is cited Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Virts, E. L. Articles by Raschke, W. C. Search for Related Content PubMed PubMed Citation Articles by Virts, E. L. Articles by Raschke, W. C.

The Role of Intron Sequences in High Level Expression from CD45 cDNA Constructs^*

Elizabeth L. Virts and William C. Raschke

From the Sidney Kimmel Cancer Center, San Diego, California 92121

Consistent expression from CD45 cDNA constructs has proven difficult to achieve. Through the use of new CD45 cDNA constructs and reporter genes, the role 5', 3', and intron sequences play in CD45 expression was determined. The CD45 polyadenylation signal sequence was fully functional in a -galactosidase reporter construct. Furthermore, the CD45 3'-untranslated region and downstream sequences were shown to contain no negative regulatory elements. Several new CD45 cDNA constructs were designed that contain either the cytomegalovirus promoter, the leukocyte function-associated antigen (LFA-1; CD11a) promoter, or various CD45 5' regions. Neither the cytomegalovirus nor the LFA-1 promoter was capable of generating detectable levels of expression in constructs with CD45 cDNA. However, when CD45 intron sequences between exons 3 and 9 were inserted in the cDNA construct to generate a CD45 minigene, the LFA-1 promoter was able to drive reproducible, significant expression of CD45. CD45 minigenes using the CD45 5' sequences up to 19 kilobases upstream of the transcriptional start produced very little protein. The LFA-1 CD45 minigene construct produced correct cell type-specific isoforms when expressed in T and B lymphocyte lines. Therefore, we conclude that the regulation of CD45 expression and cell type-specific splicing requires elements within the intron sequences.

This article has been cited by other articles:

				A. B. Rose, T. Elfersi, G. Parra, and I. Korf Promoter-Proximal Introns in Arabidopsis thaliana Are Enriched in Dispersed Signals that Elevate Gene Expression PLANT CELL, March 1, 2008; 20(3): 543 - 551. [Abstract] [Full Text] [PDF]

				M.-X. Zhang, H. Ou, Y. H. Shen, J. Wang, J. Wang, J. Coselli, and X. Li Wang Regulation of endothelial nitric oxide synthase by small RNA PNAS, November 22, 2005; 102(47): 16967 - 16972. [Abstract] [Full Text] [PDF]

				L. C. Filip and N. I. Mundy Rapid Evolution by Positive Darwinian Selection in the Extracellular Domain of the Abundant Lymphocyte Protein CD45 in Primates Mol. Biol. Evol., August 1, 2004; 21(8): 1504 - 1511. [Abstract] [Full Text] [PDF]

				E. L. Virts, O. Diago, and W. C. Raschke A CD45 minigene restores regulated isoform expression and immune function in CD45-deficient mice: therapeutic implications for human CD45-null severe combined immunodeficiency Blood, February 1, 2003; 101(3): 849 - 855. [Abstract] [Full Text] [PDF]