(
A) PCR using the genomic DNA of
5A7 and 4A+ with primers specific to
CHLI1 and four neighboring genes (
UP1,
UP2,
UP3,
UP4) downstream of the
CHLI1 gene. The sizes of the genomic DNA PCR products for
CHLI1,
UP1,
UP2,
UP3 and
UP4 are, 459, 100, 342, 550 and 672 (bp), respectively. Odd numbered lanes denote
5A7; even numbered lanes denote 4A+; ST denotes 1 kb plus DNA ladder. (
B) PCR using the genomic DNA of
5A7 and 4A+ with primers specific to
CHLI1 and four neighboring genes (
FDX3, AMT, UP5 and
UP6) upstream of the
CHLI1 gene. The sizes of the genomic DNA PCR products for
CHLI1,
FDX3, AMT, UP5 and
UP6 are, 459, 90, 369, 379 and 369 (bp), respectively. Odd numbered lanes denote
5A7; even numbered lanes denote 4A+; M denotes 50 bp DNA ladder (NEB, Beverly, MA). (
C) PCR and RT-PCR with
UP6 gene specific primers using the
5A7 and 4A+ genomic DNA and cDNA. Actin was used as a control. Actin genomic and cDNA product sizes are 527 and 305 (bp), respectively. Odd numbered lanes denote genomic DNA PCR products; even numbered lanes denote cDNA products. All primers used spanned an intron. M denotes 50 bp DNA ladder. All DNA samples were run on a 1.8% agarose gel. Gene names are given at the bottom of the gel. Primer sequences are shown in
Table 2 and
Table 3.