A high-efficiency vector for expression of foreign genes in myeloma cells.
We have constructed a high-efficiency vector for expression of genes of interest in myeloma cells. This vector is comprised of regulatory sequences from immunoglobulin (Ig) genes, including the heavy-chain enhancer, a kappa light-chain promoter and splice site, and the polyadenylation signal downstream from the kappa constant region. The expression capacity of this vector was assayed in J558L myeloma cells using human tissue plasminogen activator as a reporter gene. Stable transfectants were analyzed for protein, RNA, DNA copy number and transcription rate. Expression was compared to that of intact, transfected Ig genes and to endogenous Ig. Tissue plasminogen activator cloned into this vector was found to be expressed as efficiently as intact, transfected Ig genes, producing 1-2% of total cellular mRNA from a single copy of the gene. RNA levels and transcription rates relative to those of endogenous Ig genes were found to be about 25% and 38%, respectively. Because of its high efficiency, potential for gene amplification, and various scale-up advantages of myeloma cells, this vector-host system may yield levels of desired proteins than currently available systems.
Hendricks
M B
MB
Integrated Genetics, Inc., Framingham, MA 01701.
Banker
M J
MJ
McLaughlin
M
M
D000818:Animals; D002460:Cell Line; D004262:DNA Restriction Enzymes; D004742:Enhancer Elements, Genetic; D005796:Genes; D005803:Genes, Immunoglobulin; D005809:Genes, Regulator; D005822:Genetic Vectors; D010954:Plasmacytoma; D010957:Plasmids; D011401:Promoter Regions, Genetic; D014158:Transcription, Genetic; D014162:Transfection
D004262:DNA Restriction Enzymes