The replacement-type vector (PmlckBSNeo2.3) was introduced into D3 embryonic stem (ES) cells by electroporation to disrupt the lck locus.
Two cell lines, 59B5 (ATCC-CRL-11115) and 56B3 (ATCC-CRL-11117), were generated that were deficient for the lck gene.
These cell lines were used to produce mutant mice with germ line transmission for lck disruption.
Heterozygous mice were inbred to obtain mice homozygous for the disrupted lck gene.
A culture submitted to the ATCC as CRL-11117 in September of 1992 was found to be contaminated with mycoplasma. Progeny were cured by a 21-day treatment with BM Cycline.
The cured cell line is available as CRL-2542. The original patent deposit is available as CRL-11117.
The line should be grown on feeder layers of mitomycin C treated (0.01 mg/ml for 90 minutes) primary mouse embryonic fibroblasts or STO cells (see ATCC CRL-1503
or ATCC 56-X.2
, MITC-STO cells).