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PCR-generated cDNA library of transition-stage maize embryos: cloning and expression of calmodulin genes during early embryogenesis
Authors:Christian Breton  Annie Chaboud  Elisabeth Matthys-Rochon  Elizabeth E M Bates  J Mark Cock  Hillel Fromm  Christian Dumas
Institution:(1) Reconnaissance Cellulaire et Amélioration des Plantes-CNRS-INRA, E.N.S. Lyon, 46 allée d'Italie, 69364 Lyon Cedex 07, France;(2) Department of Plant Genetics, Weizmann Institute of Science, 76100 Rehovot, Israël;(3) Present address: Centre de Recherches Forestières, I.N.R.A. Orléans, 45160 Ardon, France
Abstract:One hundred maize zygotic embryos microdissected at the transition stage were used to construct a cDNA library after non-selective PCR (NS-PCR) amplification of whole cDNA populations. The library contains 2.3 × 105 recombinants and two different calmodulin cDNAs were cloned using a heterologous probe from petunia. Calmodulin expression was confirmed throughout maize embryogenesis at the mRNA, amplified cDNA and protein levels. Sequence analysis suggests a maize origin for both clones and negligible nucleotide changes linked to PCR. This library is the first described for early plant embryos and represents a breakthrough to isolate genes involved in embryo differentiation.
Keywords:calmodulin  cDNA library  embryogenesis  PCR  transition stage  Zea mays
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