Saltar al contenido
Merck

Dosage Compensation of X-Linked Muller Element F Genes but Not X-Linked Transgenes in the Australian Sheep Blowfly.

PloS one (2015-10-28)
Rebecca J Linger, Esther J Belikoff, Maxwell J Scott
RESUMEN

In most animals that have X and Y sex chromosomes, chromosome-wide mechanisms are used to balance X-linked gene expression in males and females. In the fly Drosophila melanogaster, the dosage compensation mechanism also generally extends to X-linked transgenes. Over 70 transgenic lines of the Australian sheep blowfly Lucilia cuprina have been made as part of an effort to develop male-only strains for a genetic control program of this major pest of sheep. All lines carry a constitutively expressed fluorescent protein marker gene. In all 12 X-linked lines, female larvae show brighter fluorescence than male larvae, suggesting the marker gene is not dosage compensated. This has been confirmed by quantitative RT-PCR for selected lines. To determine if endogenous X-linked genes are dosage compensated, we isolated 8 genes that are orthologs of genes that are on the fourth chromosome in D. melanogaster. Recent evidence suggests that the D. melanogaster fourth chromosome, or Muller element F, is the ancestral X chromosome in Diptera that has reverted to an autosome in Drosophila species. We show by quantitative PCR of male and female DNA that 6 of the 8 linkage group F genes reside on the X chromosome in L. cuprina. The other two Muller element F genes were found to be autosomal in L. cuprina, whereas two Muller element B genes were found on the same region of the X chromosome as the L. cuprina orthologs of the D. melanogaster Ephrin and gawky genes. We find that the L. cuprina X chromosome genes are equally expressed in males and females (i.e., fully dosage compensated). Thus, unlike in Drosophila, it appears that the Lucilia dosage compensation system is specific for genes endogenous to the X chromosome and cannot be co-opted by recently arrived transgenes.

MATERIALES
Número de producto
Marca
Descripción del producto

Sigma-Aldrich
Alcohol etílico puro 200, Molecular Biology
Sigma-Aldrich
Alcohol etílico puro, 200 proof, ACS reagent, ≥99.5%
Sigma-Aldrich
Cloroformo, contains 100-200 ppm amylenes as stabilizer, ≥99.5%
Sigma-Aldrich
Dodecilsulfatosódico, BioReagent, suitable for electrophoresis, Molecular Biology, ≥98.5% (GC)
Sigma-Aldrich
Alcohol etílico puro, 200 proof, meets USP testing specifications
Sigma-Aldrich
Cloroformo, contains ethanol as stabilizer, ACS reagent, ≥99.8%
Sigma-Aldrich
Dodecilsulfatosódico, ≥99.0% (GC), dust-free pellets
Sigma-Aldrich
Alcohol etílico puro 190, for molecular biology
Sigma-Aldrich
Dodecilsulfatosódico, ACS reagent, ≥99.0%
Sigma-Aldrich
Sodium dodecyl sulfate solution, BioUltra, Molecular Biology, 10% in H2O
Sigma-Aldrich
Dodecilsulfatosódico, ReagentPlus®, ≥98.5% (GC)
Sigma-Aldrich
Fenol solution, Equilibrated with 10 mM Tris HCl, pH 8.0, 1 mM EDTA, BioReagent, Molecular Biology
Sigma-Aldrich
Proteinasa K from Tritirachium album, lyophilized powder, Molecular Biology, BioUltra, ≥30 units/mg protein
Sigma-Aldrich
Fenol:cloroformo:alcohol isoamílico 25:24:1, saturado con Tris 10 mM, pH 8,0, EDTA 1 mM, Supplied with Equilibration buffer, Molecular Biology
Sigma-Aldrich
Cloroformo, contains amylenes as stabilizer, ACS reagent, ≥99.8%
Sigma-Aldrich
Fenol, puriss., meets analytical specification of Ph. Eur., BP, USP, ≥99.5% (GC), crystalline (detached)
Sigma-Aldrich
Dodecilsulfatosódico, BioUltra, Molecular Biology, ≥99.0% (GC)
Sigma-Aldrich
Sodium dodecyl sulfate solution, BioUltra, 20% in H2O
Sigma-Aldrich
Etanol, purum, absolute ethanol, denaturated with 4.8% isopropanol, A15 IPA1, ≥99.8% (based on denaturant-free substance)
Sigma-Aldrich
Cloroformo, ACS spectrophotometric grade, ≥99.8%, contains 0.5-1.0% ethanol as stabilizer
Sigma-Aldrich
Cloroformo, ReagentPlus®, ≥99.8%, contains 0.5-1.0% ethanol as stabilizer
Supelco
Dodecilsulfatosódico, dust-free pellets, suitable for electrophoresis, Molecular Biology, ≥99.0% (GC)
Sigma-Aldrich
Cloroformo, anhydrous, contains amylenes as stabilizer, ≥99%
Sigma-Aldrich
Fenol solution, Saturated with 0.01 M citrate buffer, pH 4.3 ± 0.2, BioReagent, Molecular Biology
Sigma-Aldrich
3-Methyl-1-butanol, reagent grade, 98%
Sigma-Aldrich
Cloroformo, contains ethanol as stabilizer, meets analytical specification of BP, 99-99.4% (GC)
Sigma-Aldrich
Fenol, ≥99%
Sigma-Aldrich
Etanol, purum, fine spirit, denaturated with 4.8% methanol, F25 METHYL1, ~96% (based on denaturant-free substance)
Sigma-Aldrich
Isoamyl alcohol, ≥98%, FG
Sigma-Aldrich
Fenol solution, ≥89.0%