Microsatellite DNA variants between the FVB/N and C3HeB/FeJLe and C57BL/6J mouse strains.

Abstract

The inbred mouse strain FVB/N has been widely used for the generation of transgenic animals because of their vigorous reproductive performance and because of the ease with which the male pronucleus is visualized, which facilitates DNA microinjection (Taketo et al. 1991). This strain originally arose from an outbred colony of N:GP Swiss mice at the National Institutes of Health (NIH). From this colony, two strains, HSFS/N and HSFR/N, were derived based on their sensitivity to pertussis vaccine inoculation after a challenge with histamine. In the 1970s, at the eighth inbred generation, a group of HSFS/N mice carrying the F v l b allele (for sensitivity to the B strain of Friend leukemia virus) were inbred and selected for F v l b homozygosity. This strain, designated FVB/ N, has been maintained since then without selection for virus type or vaccine sensitivity. The utility of FVB/N for genetic mapping studies is limited by the lack of information regarding DNA variant alleles between this and other inbred laboratory mouse strains. In this communication we report the identification of microsatellite sequences that are variant between FVB/N and the strains C3HeB/FeJLe and C57BL/6J. FVB/N, C3HeB/FeJLe, and C57BL/6J females were obtained from The Jackson Laboratory. Genomic DNA was prepared from mouse tails and amplified by PCR with microsatellite markers developed at the Whitehead Institute/MIT Center for Genome Research and obtained from Research Genetics (Huntsville, Ala). PCR was carried out in either of two protocols. Protocol 1 was as follows: an aliquot of 100 ng of genomic DNA was amplified in 20 I~1 PCR reaction containing 10 rnM Tris-HC1 (pH: 8.0), 50 rn~ KCI, 1 p.M of each primer, 2.0 mM MgC12, 250 I~M each dNTP, and 2.5 units of Taq polymerase (Boehringer-Mannheim, Ind.). PCR reactions were overlaid with 20 txl of mineral oil (Sigma, Mo.) and amplified in a thermal cycler (Hybaid) as follows: 1 cycle 95~ for 5 min, 60~ for 2 rain, and 72~ for 3 min; 30 cycles of 95~ for 1 min, 60~ for 2 rain, and 72~ for 3 min. A final elongation step at 72~ for 10 rain followed. Protocol 2 was as follows: an aliquot of 20 ng of genomic DNA was amplified in 20 ~1 PCR reaction containing 22 na~ Tris-HC1 (pH: 8.4), 55 nan KC1, 0.66 ~M of each primer, 1.65 mu MgC12, 880 IxM dNTPs, and 0.44 units of Taq polymerase (Gibco-BRL). PCR reactions were amplified in a thermal cycler (MJ Research) as follows: 1 cycle 95~ 30 cycles of 95~ for 45 s; 45-55~ for 45 s, and 72~ for 2 min and 30 s. A final elongation step at 72~ for 5 min followed. PCR products were run in 3% agarose gels. When necessary, PCR products were labeled in the presence of [o~-32P] dCTP (NEN), by including 0.125 Ixl of a stock solution (3000 Ci/mmol) in a 10-txl PCR reaction. Radiolabeled DNA products were run in denaturing poly-