Below can be seen some in situ hybridisation with genomic mink DNA on some of the hybrid cell lines. (The in situ hybridisation was done by Terje Raudsepp from this department). The mink material appear yellow on the hamster chromosome background. The number of mink origin 'chromosomes' vary between the hybrid cell lines. The numbers refer to the identification of the cell lines. In the picture from line 9353 can be seen an interphase cell which also show strong mink signal.
A mink primer has been constructed which preferably amplify mink DNA from the hybrid cell lines.
The construction of the primer has been done using the mink tyrosine aminotransferase
gene (AF163863 , Prieur et al. unpublished)
as blast search sequence in the NCBI nucleotide database. Five more or less
identical stretches
hit other mink sequences and the consensus mink sine primer was chosen from
the from nucleotide 5244 to 5261 as gcacctgggtggctcagt.
Results of using that mink sine primer on the RH cell panel is shown below where the
hamster only give single band, cell lines with positiv mink content give a smear and
some lines only contain the hamster bands and a few weak extra bands
which probably are linked to the tymidine kinase gene
which were the selective marker. These cell lines will be discarded as they never gave
any hits with any of the 59 markers.
Annother sine
primer was constructed 'tggcttaacccactgagc' 5272 - 5255 which goes
out of the sine sequence and therfore contains less sine material.
When the PCR product from a mink-hamster hybrid cell line was used to
paint back to mink chromosomes.
the retaind chromosome sections is seen, as shown in the picture. This hybrid cell
line number 9480 has retaind parts of the mink chromosomes 5, 8 and 11.