Dot-blot analysis of ATHCKA1
binding to cDNA inserts of other protein kinases (A) and northern-blot
analyses of cdc2 in wild type (WT) and the transformants
(D, F, G, H, and M) (B). A, Either 100 ng (1×) or 200 ng (2×) of the
1.2-kb ATHCK2A1 cDNA insert (column 3, second lane) was
loaded, and the amount of other DNAs (the number of molecules) was
adjusted according to their size. As a negative control, the cDNA
insert of CK2B1 and the plasmid 100 ng of pUC18 was
loaded. Lane 1, CK2B1 insert (1.1 kb,
SalI/XbaI cut); lane 2, pUC18; lane 3,
CK2A1 insert (1.2 kb,
SalI/XbaI cut); lane 4,
cdc2a (1.4 kb, EcoRI cut); lane 5, clone
2F1T7P Arabidopsis protein kinase homolog (0.7 kb,
SalI/XbaI cut); and lane 6,
plant-specific protein-kinase clone CD-18 (0.63 kb,
EcoRI cut). B, Northern-blot analyses of the expression
of cdc2 kinase in the wild type and the transformants.
Poly(A+) RNA was isolated from the total RNA extracted from
21-d-old light-grown plants and 0.5 μg was loaded in each lane.
Riboprobes were synthesized as described in Methods. The probed blot was stripped off using a kit and
reprobed.