research animal models

ORIGIN
At the Ludwig Institute for Cancer Research, a hybrid construct containing H-2K^b (MHC Class I antigen) 5' promotor sequences fused to the early region of the SV40 mutant tsA58, which encodes both the large and small SV40 tumor antigens, was microinjected into fertilized oocytes from CBA/Ca x C57BL/10 F1 mice. Following reimplantation, 88 mice were born, of which 34 were transgenic and carried one to five copies of the gene. RNA from a variety of tissues from one nontransgenic and three transgenic animals was analyzed by Northern blot analysis using a SV40 early region-specific probe. RNA extracted from tissues of transgenic mice contained varying amounts of a 2.5kb RNA species, while no tsA58TAg RNA was detected in tissues of the nontransgenic mouse; thymus and liver showed the highest level of expression, while brain showed the lowest. Distribution rights to Charles River in 1991.

STRAIN CODE
237 (HOMOZYGOUS)
238 (HETEROZYGOUS)

COAT COLOR
Primarily agouti, infrequently black

TRANSGENE INFORMATION
Construction of this transgene involved fusion of the 5'-flanking promoter sequences as well as the transcriptional initiation site of the H-2K^b Class 1 gene to the tsA58 early region coding sequences. The 4.2kb EcoRI to NruI fragment encompassing the H-2K^b promoter sequences was ligated to the 2.7kb BglI to BamHI fragment derived from tsA58 early region and pUC19 double digested with EcoRI and BamHI. The Bg1I site had been blunted using the Klenow fragment of E. coli DNA Polymerase1 to allow fusion to the Nru1 site. For purposes of microinjection, the H-2K^btsA58 DNA fragment was isolated free of vector sequences by digestion with EcoRI and SalI.

RESEARCH APPLICATION
oncology

ORDERING INFORMATION
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