Molecular characterization of OsPAP2: transgenic expression of a purple acid phosphatase up-regulated in phosphate-deprived rice suspension cells.
A phosphate starvation-induced, purple, acid phosphatase cDNA was cloned from rice, Oryza sativa. The cDNA encoding the phosphatase (OsPAP2) has 1,893 bp with an open reading frame of 630 amino acid residues. The deduced amino acid sequence of OsPAP2 shows identities of 60-63% with other plant purple acid phosphatases and appears to have five conserved motifs containing the residues involved in metal binding. OsPAP2 expression is up-regulated in the rice plant and in cell cultures in the absence of phosphate (P( i )). The induced expression of OsPAP2 is a specific response to P( i ) starvation, and is not affected by the deprivation of other nutrients. OsPAP2 expression was responsive to the level of P( i )-supply, and transcripts of OsPAP2 were abundant in P( i )-deprived roots. The OsPAP2 cDNA was expressed as a 69 kDa polypeptide in baculovirus-infected insect Sf9 cells. In addition, the OsPAP2 gene was introduced into Arabidopsis via an Agrobacterium-mediated transformation. Functional expression of the OsPAP2 gene in the transgenic Arabidopsis line was confirmed by northern and western blot analyses, as well as by phosphatase activity assays. These results suggest that the OsPAP2 gene can be used to develop new transgenic dicotyledonous plants that are able to adapt to P( i )-deficient conditions.
Hur
Yeon Jae
YJ
College of Natural Resources and Life Science, Dong-A University, Busan, 604-714, Korea.
Jin
Byung Rae
BR
Nam
Jaesung
J
Chung
Young Soo
YS
Lee
Jai Heon
JH
Choi
Hong Kyu
HK
Yun
Dae Jin
DJ
Yi
Gihwan
G
Kim
Yong Hwan
YH
Kim
Doh Hoon
DH
D000135:Acid Phosphatase / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D000595:Amino Acid Sequence; D000818:Animals; D017360:Arabidopsis / Q000201:enzymology / Q000235:genetics / Q000378:metabolism; D015152:Blotting, Northern; D002460:Cell Line; D002478:Cells, Cultured; D018506:Gene Expression Regulation, Plant; D006023:Glycoproteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D008969:Molecular Sequence Data; D012275:Oryza / Q000201:enzymology* / Q000235:genetics; D010710:Phosphates / Q000172:deficiency* / Q000502:physiology; D010940:Plant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D030821:Plants, Genetically Modified / Q000201:enzymology / Q000235:genetics / Q000378:metabolism; D016133:Polymerase Chain Reaction; D012231:Rhizobium / Q000235:genetics; D017386:Sequence Homology, Amino Acid; D018411:Spodoptera
D006023:Glycoproteins; D010710:Phosphates; D010940:Plant Proteins; C091137:purple acid phosphatase; D000135:Acid Phosphatase