By mRNA differential display from control versus NaCl_shocked Arabidopsis seedlings, we screened an Arabidopsis 3′ partial cDNA, which represents a gene encoding inositol 1,3,4_trisphosphate (Ins(1,3,4)P 3) 5/6_kinase_like protein. Northern blotting analysis showed that the gene, named as AtITL1, is strongly induced by NaCl and low temperature, but not induced by drought and abscisic acid (ABA). Analysis of 5′ region of the AtITL1 found that there are dehydration_responsive element/C_repeat (DRE/CRT) cis _acting elements, but no elements related to G_box and ABRE (ABA_responsive element) in its 5′ region, which is consistent with the expression patterns of the AtITL1 independent of ABA. These results suggest that the AtITL1 may be involved in the osmotic stress response pathway independent of ABA.
Protein phosphorylation and dephosphorylation are the general means of regulation metabolism within a cell. A PKA catalytic subunit was found in Arabidopsis genome using Blast software. The cDNA was cloned by RT-PCR and sequencing result indicated a high degree of homology at protein level. The cDNA was subcloned into pET30a (+) and expressed in E. coli at different temperatures. The target protein was insoluble when induced at 37degreesC, while dissolvable if induced at 22degreesC with 0.01 mmol/L IPTG. Ni2+-NTA affinity chromatography was used to purify the target protein, which was shown to have cAMP-dependent protein kinase activity. Western blotting analysis indicated that stress treatments affected the expression of PKA catalytic subunit at protein level just to a small extent.