Indeed, the angora mouse mutant carries a 2-kb loss-of-function deletion in Fgf5 and displays a delayed catagen onset and excessively long truncal hair [ 7].
To generate H. volcanii insertion mutants, a 4-6 kb fragment mixture was isolated from the plasmid library, and the mixture was used to transform H295 cells for chromosomal targeting en masse, in which each fragment would target its counterpart genomic locus.
In the ΔIS481-BP155 mutant, a 2.2-kb insertion containing an intact bteA 5'-UTR (derived from B. pertussis Tohama) replaced the native bteA 5'-UTR + IS481a gene.
In the suspected mutant, a 3.6 kb restriction fragment hybridized with the SpcR probe (Fig. 3B, lane 2), while no hybridization signal was detected when the internal fragment of salKR and pUC18 served as the probes (Fig. 3C and 3D, lane 2).
For the construction of a toaD mutant, a 2 kb fragment carrying the toaD gene was cloned into pJQ200SK [ 103] and a spectinomycin resistance cassette ligated into the unique NcoI site within toaD.
Mouse models of Stz1 mutants, harboring a 300 kb deletion on mouse chromosome 2 including Kcnq2, Arfgap1, and Chrna4, demonstrate an increased susceptibility to seizures [16].
Moreover, Southern blot showed that a single 4.4 kb band with expected size was identified in all nine Mosdi1 ReGFP mutants while an 8.8 kb band was found in wild type (Fig. 2d, f).
This type of mutation is not typical for EMS mutagenesis; however, our previous studies of mutant pro41 (also a product of EMS mutagenesis) showed that this mutant carries a 4-kb deletion that includes the pro41 ORF (Nowrousian et al. 2007).
In contrast, +IS481-BP157 mutant had a 3.2-kb insertion containing a bteA 5'-UTR + IS481a (derived from B. pertussis BP155) instead of its own bteA 5'-UTR.
The InvA mutant carries a 18 kb inversion encompassing the rrnA operon.
