PJB-2019-220
Cloning and transcriptional analysis of RuBisCO hybrid promoter
Ammara Masood, Hira Mubeen, Nadia Iqbal and Aftab Bashir
Abstract
Two variants of RuBisCO promoter from zea mays were selected to generate hybrid promoter by using bioinformatics tools. Sequence analysis of both RuBisCO promoter variants revealed several critical cis regulatory elements and transcription factors binding sites within the promoter region. Various regulatory motifs related to constitutive expression were located in RuBisCO promoter fragments. Analysis of cis-regulatory regions has paved way to design synthetic promoters. Both variants were separately cloned in TA vector (pTZ57R/T) and then joined to get the complete hybrid promoter (Rub-H). Hybrid RuBisCO promoter was further cloned in expression vector pGR1. Transient GUS assay revealed that hybrid promoter exhibited endosperm specific expression in wheat. From the study it is demonstrated that hybrid promoter (Rub-H) may be used to derive constitutive expression in monocots. The present work provides an important insight in the designing of hybrid monocot promoters to improve various traits in crops without facing IPR issues. It is expected that complete understanding of the regulatory regions and transcription factors in the regulatory regions would help in designing new synthetic/ hybrid promoters for tissue specific or constitutive expression of transgenes
To Cite this article:
Masood, A., H. Mubeen, N. Iqbal and A. Bashir. 2021. Cloning and transcriptional analysis of RuBisCO hybrid promoter. Pak. J. Bot., 53(3): DOI: http://dx.doi.org/10.30848/PJB2021-3(27)
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