CKI1, a Histidine Kinase Homolog Implicated in Cytokinin Signal Transduction

These results suggest that CKI1 is involved in cytokinin signal transduction, possibly as a cytokinin receptor

Tatsuo Kakimoto

2002

Scholarcy highlights

  • Cytokinin plays a central role in plant development, little is known about cytokinin signal transduction
  • DNA isolated from the cki line was digested with Spe I, self-ligated, and used to transform Escherichia coli by electroporation, and the plasmid was purified from the ampicillin-resistant E. coli cells as described elsewhere [; A Spe I digest of pC1S1 was ligated to the Xba I fragment of pPCVICEn4HPT, which carries the T-DNA right border sequence, the vegetative, the conjugational DNA replication origins of plasmid RK2, and the T-DNA left border sequence, to yield Ti plasmid pC1S1Ti
  • The amplified DNA was used to screen a cDNA library that had been constructed in Lambda ZAP II, in accordance with the manufacturer's instructions, with polyadenylated RNA from shoot tissue of the cki line
  • For construction of the Ti plasmid carrying CKI1 cDNA linked to the CaMV 35, S RNA promoter, 157 bp of the 5′ noncoding sequence of cCKI1-16 was removed by treatment with exonuclease III, and all of the 3′ noncoding sequence was removed by digestion with Spe I
  • B., Methods Enzymol.153 253] next to the CaMV 35, S RNA promoter, and the plasmid that contained the insert in the sense orientation was selected and designated p35ScCKI1
  • Walden for providing plasmid pPCVICEn4HPT, for comments on the manuscript, and for continuous encouragement

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