Derivation and partial analysis of two highly active myeloma cell transfectants.
Vectors have been designed to optimise the expression of heterologous proteins in transfected mouse myeloma cells. The over-ridingly important DNA element contained in these constructs is the classical mouse immunoglobulin heavy chain enhancer. It is shown that even in the absence of a well-known promoter element, the enhancer can drive gene expression in stable cell transfectants and the main transcriptional start site utilized in such situations has been mapped to within the previously defined enhancer region. Using chicken lysozyme as a reporter function in these vectors, two transfected myeloma cell clones have been isolated which secrete this protein at levels 50-100-times as high as those usually obtained with the same vectors and it is shown that in molar terms this is at least as high as endogenous immunoglobulin produced by a related line. Analysis of these lines show that in one case only a single copy, and in the other two to three copies, of the apparently unrearranged vector have integrated at a single locus within the genome. Possible explanations for the high-level expression are discussed.
Department of Biochemistry, University of Leicester, UK.
T M
TM
Harrison
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K
Hudson
S E
SE
Munson
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S
Uff
S
S
Glassford
D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D002645:Chickens; D005822:Genetic Vectors; D007074:Immunoglobulin G / Q000032:analysis*; D051379:Mice; D008969:Molecular Sequence Data; D009113:Muramidase / Q000032:analysis*; D010957:Plasmids; D014162:Transfection
D007074:Immunoglobulin G; D009113:Muramidase