Koizumi Nozomu, Toyota Kentaro, Kitajima Sakihito, Yamada Yasuyuki, Sato Fumihiko
Division of Applied Life Sciences, Graduate School of Agriculture, Kyoto University, Sakyo, Japan.
Biosci Biotechnol Biochem. 2002 Aug;66(8):1691-6. doi: 10.1271/bbb.66.1691.
Three genes encoding phosphoenolpyruvate carboxylase were isolated from Nicotiana sylvestris and designated Nsppc1-3. Sequencing of nucleotides showed that the coding sequence and deduced amino acid sequences were highly conserved among the genes, but sequences for noncoding regions including introns and 5'-flanking regions were not conserved. Analysis of the transcript level of the genes by a combination of reverse transcriptase-PCR and restriction fragment polymorphism showed mostNsppc1 in the leaves, stems, roots, and cultured cells of N. sylvestris. beta-Glucuronidase activity was detected histochemically in mesophyll cells in leaves, lateral buds, and vascular bundles in roots of transgenic tobacco harboring a chimeric construct of the Nsppc1 promoter and the gene for beta-glucuronidase. Deletion analysis indicated the presence of a silencer-like element for basal expression in the promoter region of Nsppc1.