Chromosome assignment of Cd36 transgenes in two rat SHR lines by FISH and linkage mapping of transgenic insert in the SHR-TG19 line.
The chromosome position of the Cd36 insert was determined by FISH in two rat transgenic lines (SHR/Ola-TgN(EF1aCd36)10Ipcv (SHR-TG10) and SHR/Ola-TgN(EF1aCd36)19Ipcv (SHR-TG19). The Cd36 transgene construct labelled with digoxigenin-11-dNTP was used as a probe in the FISH analysis. In accord with the previous finding that the SHR-TG10 harbours 6-8 copies of the transgene, the signals from both metaphase and interphase nuclei of SHR-TG10 preparations were rather strong and the probe hybridized to both copies of chromosome 1 at band q55. The probe hybridization to SHR-TG19 metaphase preparations also showed homozygosity of the transgene with localization of both copies to chromosome 11 at band q11. The signals were distinct but much weaker compared to the SHR-TG10, which again is in accord with the fact that the SHR-TG19 line harbours only a single copy of the transgene. In order to look for a possible impact of the insertion site neighbourhood upon the transgene phenotypic effect, we performed linkage mapping of the transgene in the SHR-TG19 line. By linkage mapping, the placement of the transgene to the proximal part of RNO11 was confirmed, the critical interval being 4 cM between D11Rat20 and D11Rat21, in good agreement with the RH map. Within the close neighbourhood of the inserted Cd36 transgene, there are several genes known to be expressed in kidney, and so the influence of some regulatory sequences enhancing kidney expression of the Cd36 transgene can be envisaged.
Institute of Biology and Medical Genetics, 1st Medical Faculty, Charles University, Albertov 4, Prague 2, 128 00, Czech Republic.
F
F
Liska
G
G
Levan
K
K
Helou
M
M
Sladká
M
M
Pravenec
V
V
Zídek
V
V
Landa
V
V
Kren
D000818:Animals; D018955:CD36 Antigens / Q000235:genetics*; D002874:Chromosome Mapping; D033481:Chromosomes, Mammalian; D020224:Expressed Sequence Tags; D017404:In Situ Hybridization, Fluorescence; D051379:Mice; D030781:Organisms, Genetically Modified; D051381:Rats; D011918:Rats, Inbred SHR; D014018:Tissue Distribution; D019076:Transgenes
D018955:CD36 Antigens