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Evaluation of novel control elements by construction of eukaryotic expression vectors

In English

Författare

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.

Publiceringsår

1997-04-01

Språk

Engelska

Sidor

191-198

Publikation/Tidskrift/Serie

Gene

Volym

188

Issue

2

Dokumenttyp

Artikel i tidskrift

Förlag

Elsevier

Nyckelord

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

Status

Published

ISBN/ISSN/Övrigt

  • ISSN: 0378-1119