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Plant Physiol, August 2001, Vol. 126, pp. 1630-1636
Sequence Architecture Downstream of the Initiator Codon Enhances
Gene Expression and Protein Stability in Plants1
Samir V.
Sawant,
Kanti
Kiran,
Pradhyumna K.
Singh, and
Rakesh
Tuli*
National Botanical Research Institute, Rana Pratap Marg,
Lucknow-226001, India
Nucleotide positions conserved on the 3' side of the initiator
codon ATG and the corresponding N-terminal amino acid residues in a
number of highly abundant plant proteins were identified by
computational analysis of a dataset of highly expressed plant genes.
The reporter genes uidA and gfp were
modified to introduce these features. Insertion of GCT TCC TCC after
the initiator codon ATG augmented expression for both the reporter
genes. The insertion of each successive codon improved the expression
of  -glucuronidase (GUS) in an incremental fashion in transient
transformation of tobacco (Nicotiana tabacum) leaves.
The insertion of alanine-serine (Ser)-Ser resulted in about a 2-fold
increase in the stability of GUS. However, this did not account for the
30- to 40-fold increase in GUS activity between the constructs coding
for methionine-alanine-Ser-Ser-GUS and the native enzyme. Substitution
of the codon for Ser at the third amino acid residue with synonymous
codons reduced GUS expression. The results suggest a role for the
conserved nucleotides in the +4 to +11 region in augmenting
posttranscriptional events in the expression of genes in plants.
1
This work was supported by fellowships (to
S.V.S. and P.K.S.) and by a research grant from The Council of
Scientific and Industrial Research, Government of India.
*
Corresponding author; e-mail rakeshtuli{at}hotmail.com; fax
0522-205836.
© 2001 American Society of Plant Physiologists
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