Kinetoplast minicircle DNA from Leishmania (Viannia) lainsoni

To investigate the phenomenon that PCR of Leishmania (V.) lainsoni minicirc les using primers B1 and B2 gives anomalous small-sized products, unlike al l other members of the Leishmania Viannia subgenus, cloned kDNA minicircles from L. (Viannia) lainsoni were sequenced using fluorescent dye terminator reactions. The sequence of L. (V.) lainsoni where the primer B2 would be e xpected to bind, was different from the other members of the L. Viannia sub genus, matching in only 7 out of 19 bases with the sequence of L. ( V.) bra ziliensis at this position. The sequence obtained from the cloned minicircl es enabled the design of a new primer which, when combined with B1, allowed the amplification of full sized minicircles in L. (V.) lainsoni, but not o ther members of the L. Viannia subgenus. Comparison of the sequence obtaine d for Leishmania (V.) lainsoni with other Leishmania minicircle DNA confirm s that Leishmania (V.) lainsoni is more similar to members of the L. Vianni a subgenus than to other Leishmania, but is distinctly different.