Generation of the MeCP2 A140V "knock-in" mouse. (1A) Diagram showing the WT Mecp2 locus, the A140V targeting vector, and the recombined MeCP2 A140V locus used to create the knock-in mouse. (1B) PCR demonstrating ES cell DNA positive (177U9, W7, and A6) and negative (G3) for homologous recombination at the 3' end of the targeting vector. Lanes "WT" and "Mut" are plasmid DNA controls demonstrating the WT and A140V mutant PCR fragments (Upper Panel). The PCR fragments were digested with Acl I which cuts only the mutant PCR fragments, confirming the presence of the A140V mutation and producing a double band (1B Lower Panel). (1C) Southern blot analysis Bam HI digested ES cell DNA. A 5.5 kilobase pair (kbp) band was detected confirming homologous recombination at the 5' end of the targeting vector. The ladder on the left is in kbp. (1D) PCR genotyping results of F1 generation females. DNA from a WT female produces a single band while DNA from a heterozygous female (Het) produces two bands. The WT and Mut lanes show control plasmid reactions.