Isolation and characterization of the promoter sequence of a cassava gene coding for Pt2L4, a glutamic acid-rich protein differentially expressed in storage roots.
Cassava is one of the most important tropical food crops for more than 600 million people worldwide. Transgenic technologies can be useful for increasing its nutritional value and its resistance to viral diseases and insect pests. However, tissue-specific promoters that guarantee correct expression of transgenes would be necessary. We used inverse polymerase chain reaction to isolate a promoter sequence of the Mec1 gene coding for Pt2L4, a glutamic acid-rich protein differentially expressed in cassava storage roots. In silico analysis revealed putative cis-acting regulatory elements within this promoter sequence, including root-specific elements that may be required for its expression in vascular tissues. Transient expression experiments showed that the Mec1 promoter is functional, since this sequence was able to drive GUS expression in bean embryonic axes. Results from our computational analysis can serve as a guide for functional experiments to identify regions with tissue-specific Mec1 promoter activity. The DNA sequence that we identified is a new promoter that could be a candidate for genetic engineering of cassava roots.
Laboratório de Biologia Molecular, Universidade Federal do Pará, Belém, PA, Brazil. bsouza@ufpa.br
C R
CR
de Souza
F J
FJ
Aragão
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EC
Moreira
C N M
CN
Costa
S B
SB
Nascimento
L J
LJ
Carvalho
D001483:Base Sequence; D001665:Binding Sites; D018076:DNA, Complementary / Q000378:metabolism; D018744:DNA, Plant / Q000378:metabolism; D017343:Genes, Plant; D018698:Glutamic Acid / Q000032:analysis; D002365:Manihot / Q000235:genetics* / Q000378:metabolism; D008969:Molecular Sequence Data; D010940:Plant Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D018517:Plant Roots / Q000235:genetics / Q000378:metabolism*; D016133:Polymerase Chain Reaction; D011401:Promoter Regions, Genetic; D017422:Sequence Analysis, DNA
D018076:DNA, Complementary; D018744:DNA, Plant; D010940:Plant Proteins; D018698:Glutamic Acid