Expression and chloroplast-targeting of active phosphoenolpyruvate synthetase from Escherichia coli in Solanum tuberosum
Panstruga R, HippeSanwald S, Lee YK, Lataster M, Lipka V, Fischer R, Liao YC, Hausler RE, Kreuzaler F, Hirsch HJ
PLANT SCIENCE

127: (2) 191-205 SEP 12 1997


Abstract:
Solanum tuberosum was transformed with a chimeric gene consisting of the constitutive 35S CaMV promoter, a potato ribulose bisphosphate carboxylase (Rubisco) small subunit transit sequence (rbcS) and the phosphoenolpyruvate synthetase gene (ppsA) from E. coli. Transgenic plants were regenerated producing (phosphoenolpyruvate-) PEP-synthetase in amounts as much as 0.1% of total soluble protein. Western blot analysis indicated that most of the protein is located in the chloroplasts and that the transit sequence is cleaved off. Electron microscopy of leaves revealed that PEP-synthetase specific immunogold labeling was most pronounced over the chloroplast matrix occurring in between the thylakoid stacks. PEP-synthetase activity was detected in isolated chloroplasts of transgenic plants. Chloroplast morphology and starch production in leaves were affected. (C) 1997 Elsevier Science ireland Ltd.

Author Keywords:
chloroplast transit sequence, immunogold labeling, LT-embedding, phosphoenolpyruvate synthetase, pyruvate, P-i dikinase, transgenic potato

KeyWords Plus:
ORTHO-PHOSPHATE DIKINASE, GENE, LEAVES, TRANSFORMATION, CARBOXYKINASE, LOCALIZATION, VECTOR, MAIZE

Addresses:
RHEIN WESTFAL TH AACHEN, INST BIOL BOT MOL GENET 1, D-52074 AACHEN, GERMANY.
CHRISTIAN ALBRECHTS UNIV KIEL, INST PHYTOPATHOL, D-24118 KIEL, GERMANY.
KOREA UNIV, COLL NAT RESOURCES, DEPT AGR BIOL, SEOUL 136011, SOUTH KOREA.
UNIV COLOGNE, INST BOT, D-50931 COLOGNE, GERMANY.

Publisher:
ELSEVIER SCI IRELAND LTD, CLARE

IDS Number:
XR178

ISSN:
0168-9452