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Carl Borrebaeck

Carl Borrebaeck

Professor

Carl Borrebaeck

Evaluation of novel control elements by construction of eukaryotic expression vectors

Author

  • Christina Furebring
  • Mats Ohlin
  • Sven Pettersson
  • Carl A K Borrebaeck

Summary, in English

A novel mammalian eukaryotic expression vector for the production of immunoglobulin heavy chain (IgH) genes has been designed. This expression vector contains the variable heavy chain (VH) promoter, the IgH intron enhancer (μE) and the IgH 3' enhancer (3'E). This construct, designated pTIF-1, was stably transfected into the myeloma cell line J558L. A fivefold increase in the expression level of a rearranged IgH gene was observed when using the pTIF-1 vector containing the 3'E compared to an expression vector lacking this enhancer. Interestingly, this positive effect on the expression level of the 3' enhancer appears to be position independent. The introduction of two recently identified Ig control elements, HS3 and HS4, to the vector cassette did not further elevate the expression level in the cell line tested. The pTIF-1 vector can be used for expression of any antibody specificity, using PCR amplification of the VDJ region of interest. Furthermore, the constant region can easily be exchanged, which further facilitates studies to dissect different effector functions of IgH constant genes.

Department/s

  • Department of Immunotechnology

Publishing year

1997-04-01

Language

English

Pages

191-198

Publication/Series

Gene

Volume

188

Issue

2

Document type

Journal article

Publisher

Elsevier

Keywords

  • 3' Enhancer
  • Antibody
  • Hypersensitive sites 3 and 4
  • Intron enhancer

Status

Published

ISBN/ISSN/Other

  • ISSN: 0378-1119