Amplification of adenosine deaminase genes in mammalian cells us

Details Amplification of adenosine deaminase genes in mammalian cells us Amplification of adenosine deaminase genes in mammalian cells us

1984-07-05

1989-04-18

Wiseman, Thomas G.

Chemistry: molecular biology and microbiology

Micro-organism, tissue cell culture or enzyme using process...

Preparing compound containing saccharide radical

435 68, 435 70, 4351723, 4353171, 435320, 935 62, 935 70, 536 27, C12P2193.4, C12P 2100, C12N 1500

Patent

active

048227366

ABSTRACT:
Disclosed are cultured mouse cells with amplified adenosine deaminase genes which greatly overproduce adenosine deaminase. The cells were isolated by growth in media containing alanosine, uridine, and a cytotoxic concentration of adenosine. Maximum enhancement of adenosine deaminase activity was achieved by also adding deoxycoformycin to the media and then serially increasing the deoxycoformycin concentration at 4 to 6 week intervals. Also disclosed is a cDNA homologous to mouse adenosine deaminase mRNA. The cDNA was produced by synthesis against RNA extracted from the cells containing amplified adenosine deaminase genes and then inserted into a bacterial plasmid. Multiple copies of the cDNA were produced by replication of the bacteria into which the plasmids were inserted.

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Yeung, C.-Y. et al., Jul. 1983, J.B.C. 258:8330.
Kaufman, R. et al., Apr. 1983, M.C.B. 3:699.

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