TY - JOUR
T1 - High-frequency transformation of human repair-deficient cell lines by an Epstein-Barr virus-based cDNA expression vector
AU - Peterson, Carolyn
AU - Legerski, Randy
N1 - Copyright:
Copyright 2014 Elsevier B.V., All rights reserved.
PY - 1991/11/15
Y1 - 1991/11/15
N2 - We constructed a human cDNA expression vector by combining an episomal Epstein-Barr virus (EBV) vector with the expression cassette from the transient-expression vector, pCDM8. This new vector, designated pEBS7, exhibited high-level expression of reporter genes in normal and repair-deficient xeroderma pigmentosum cell lines. Reconstruction experiments indicated that marker genes diluted to a frequency of 10-5 can be rescued on a single transfection dish. Moreover, derivative cell lines that constitutively express the gene encoding EBV nuclear antigen 1 exhibited a tenfold enhancement in the frequency of rescue of marker genes. The feasibility of preparing large-scale directional or nondirectional cDNA libraries in pEBS7 was demonstrated and reconstruction experiments indicated that marker genes could be rescued from either library with equal efficiency. These results establish a high-efficiency system for the isolation of genes by direct phenotypic selection in human mutant cell lines.
AB - We constructed a human cDNA expression vector by combining an episomal Epstein-Barr virus (EBV) vector with the expression cassette from the transient-expression vector, pCDM8. This new vector, designated pEBS7, exhibited high-level expression of reporter genes in normal and repair-deficient xeroderma pigmentosum cell lines. Reconstruction experiments indicated that marker genes diluted to a frequency of 10-5 can be rescued on a single transfection dish. Moreover, derivative cell lines that constitutively express the gene encoding EBV nuclear antigen 1 exhibited a tenfold enhancement in the frequency of rescue of marker genes. The feasibility of preparing large-scale directional or nondirectional cDNA libraries in pEBS7 was demonstrated and reconstruction experiments indicated that marker genes could be rescued from either library with equal efficiency. These results establish a high-efficiency system for the isolation of genes by direct phenotypic selection in human mutant cell lines.
KW - DNA transfection
KW - EBV nuclear antigen 1
KW - Episomal vector
KW - expression libraries
KW - repair-deficient mutants
KW - reporter genes
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U2 - 10.1016/0378-1119(91)90328-9
DO - 10.1016/0378-1119(91)90328-9
M3 - Article
C2 - 1660831
AN - SCOPUS:0025935425
SN - 0378-1119
VL - 107
SP - 279
EP - 284
JO - Gene
JF - Gene
IS - 2
ER -