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Evaluation of novel control elements by construction of eukaryotic expression vectors

Furebring, Christina LU ; Ohlin, Mats LU ; Pettersson, Sven and Borrebaeck, Carl A K LU (1997) In Gene 188(2). p.191-198
Abstract

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... (More)

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.

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Please use this url to cite or link to this publication:
author
organization
publishing date
type
Contribution to journal
publication status
published
keywords
3' Enhancer, Antibody, Hypersensitive sites 3 and 4, Intron enhancer
in
Gene
volume
188
issue
2
pages
8 pages
publisher
Elsevier
external identifiers
  • Scopus:0030939563
ISSN
0378-1119
DOI
10.1016/S0378-1119(96)00791-3
language
English
LU publication?
yes
id
50bdd625-aa6e-416e-ac3f-eb60fbd4bff5
date added to LUP
2016-04-19 14:07:59
date last changed
2016-05-17 10:49:42
@misc{50bdd625-aa6e-416e-ac3f-eb60fbd4bff5,
  abstract     = {<p>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.</p>},
  author       = {Furebring, Christina and Ohlin, Mats and Pettersson, Sven and Borrebaeck, Carl A K},
  issn         = {0378-1119},
  keyword      = {3' Enhancer,Antibody,Hypersensitive sites 3 and 4,Intron enhancer},
  language     = {eng},
  month        = {04},
  number       = {2},
  pages        = {191--198},
  publisher    = {ARRAY(0x9cc75d0)},
  series       = {Gene},
  title        = {Evaluation of novel control elements by construction of eukaryotic expression vectors},
  url          = {http://dx.doi.org/10.1016/S0378-1119(96)00791-3},
  volume       = {188},
  year         = {1997},
}