Analysis of polarity in the expression from a multifactorial bidirectional promoter designed for high-level expression of transgenes in plants.
A synthetic bidirectional expression module was constructed by placing a computationally designed minimal promoter sequence on the 5' and 3' sides of a transcription activation module. The activation of transcription from the unidirectional and bidirectional promoters constructed from the same sequence elements was evaluated by using the reporter genes gusA and gfp. The analysis based on transient and stable transformation of tobacco showed that the artificially designed multifactorial activation module activated transcription simultaneously to comparable levels in both the directions. The transcription activation module responded to elicitors like salicylic acid, NaCl and IAA in the forward as well as reverse directions. The concentration of the elicitor required for highest gene activation was similar for the two directions in case of the three activators. The kinetics of time of induction was similar in the two directions for salicylic acid and NaCl. In the case of IAA, the transcription activation was faster in the reverse direction. The results show that constitutive and chemically inducible bidirectional promoters can be deployed for predictable simultaneous regulation of two genes for genetic engineering in plants.
Chaturvedi
Chandra Prakash
CP
National Botanical Research Institute, Rana Pratap Marg, Lucknow 226001, Uttar Pradesh, India.
Sawant
Samir V
SV
Kiran
Kanti
K
Mehrotra
Rajesh
R
Lodhi
Niraj
N
Ansari
Suraiya Anjum
SA
Tuli
Rakesh
R
D005163:Factor Analysis, Statistical; D015870:Gene Expression / Q000502:physiology*; D018506:Gene Expression Regulation, Plant / Q000235:genetics; D024861:Genetic Enhancement / Q000379:methods; D010940:Plant Proteins / Q000235:genetics* / Q000378:metabolism; D030821:Plants, Genetically Modified / Q000378:metabolism*; D011401:Promoter Regions, Genetic / Q000235:genetics*; D015202:Protein Engineering / Q000379:methods*; D017422:Sequence Analysis, DNA / Q000379:methods; D014026:Nicotiana / Q000235:genetics* / Q000378:metabolism; D015533:Transcriptional Activation; D019076:Transgenes / Q000235:genetics*
D010940:Plant Proteins