Two rounds of PCR were performed to amplify a 580-bp region of the murine Ramp2 gene using crude DNA as template.
Manual examination of the expression of these genes using human and murine gene expression databases (GEO database; GDS182 (mouse) and GDS1402 (human); [ 18]) generally revealed ubiquitous expression patterns but with bias towards greater expression in epithelia.
In addition, we are disrupting targeted virulence genes using biolistic transformation and assessing their role in the virulence of C neoformans in rabbit and murine models of cryptococcal disease.
To generate pMSCV-ΔNp63 α, a PCR fragment containing the mouse ΔNp63α gene using murine pcDNA3-ΔNp63 α as a template was subcloned into the retroviral vector pMSCV-puro (Clontech, Palo Alto, CA, USA) digested with BglII and HpaI.
A learning deficit was also observed in murine models overexpressing the DYRK1A gene, using the Morris water maze with direct and reverse platform paradigms.
Enrichment test for k-mers in 3'-UTR of murine genes was carried out using the AMADEUS tool [ 21].
A 2-fragment multiplex PCR assay was performed to amplify a 154 bp transgene sequence, using primers TransF[6FAM] and TransR (Table 1) and a 201 bp fragment belonging to the murine β-actin gene, used as a normalization control, by using primers MuActF1[6FAM] and MuActR1 (Table 1).
Real time PCR was performed with murine gene specific primers using an Applied Biosystems 7900HT Fast Real Time PCR System (Applied Biosystems, Foster City, CA).
For example, genetic ablation of the murine gene encoding TGFβR2 (Tgfbr2) using Tie1-Cre leads to lethality by E10.5 resulting from heart and yolk sac vascular defects (Carvalho et al., 2007).
The Affymetrix Mouse Genome 430 2.0 array, representing 39 000 murine gene transcripts, was used to transcriptionally profile DN3 pre-T cells purified from the thymi of CD3 C3 transgenic mice.
