Molecular characterisation
of the LdTOP2 locus. (A) Southern
blot showing that the LdTOP2 is a single copy gene.
Lanes 1–5, restriction digests of L.donovani D1700
genomic DNA with SalI, HindIII, PstI, EcoRV and EcoRV–XbaI, respectively, and their hybridisations with
the 800 bp gene internal probe of LdTOP2 corresponding
to positions 1537–2335. The positions of DNA molecular
weight markers are indicated on the left of the figure. (B)
Chromosomal location of the LdTOP2 gene. a, Chromosomes
of S.cerevisiae and L.donovani D1700
separated and visualised with ethidium bromide before blotting;
b, the same blots probed with yeast genomic DNA for S.cerevisiae and LdTOP2 gene probe for L.donovani.
Arrow shows the hybridising band of L.donovani.
PFGE running conditions as follows: initial switch time, 1800 s;
final switch time, 1800 s; run time, 72 h; current, 2 V/cm;
including angle, 106°; actual current,
30 mAmp.