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. 1998 Mar 3;95(5):2597–2602. doi: 10.1073/pnas.95.5.2597

Figure 1.

Figure 1

Strategy for targeted replacement of the N-CAM allele with lacZ and analysis of N-CAM expression in knockout mice. (A) Diagram of the N-CAM locus in heterozygous animals. In heterozygous mice, N-CAM promoter activity can be assayed both by β-gal activity and by quantitating N-CAM mRNA. (B) Structures of the N-CAM targeting vector, wild-type N-CAM allele, and disrupted N-CAM allele. Restriction enzyme sites are abbreviated as follows: E, EcoRI; K, KpnI; N, NotI; S, SalI; Sa, SacII; X, XhoI. (C) Northern blot. Total RNA isolated from the brains of wild-type (+/+), heterozygous N-CAM knockout (+/−), and N-CAM knockout (−/−) mice was hybridized to a 300-bp probe for N-CAM. (D) Ethidium bromide stain of the Northern gel shows approximately equal loading of the RNA. (E) Western blot of 30 μg of total protein isolated from the brains of wild-type (+/+), heterozygous (+/−), and homozygous (−/−) N-CAM knockout mice with a polyclonal antibody to N-CAM.