Molecular Biology Reports

, Volume 35, Issue 2, pp 81–88 | Cite as

Temporal and spatial expression analysis of gamma kafirin promoter from Sorghum (Sorghum bicolor L. moench) var. M 35-1

  • Avinash Mishra
  • Akash Tomar
  • Sangita Bansal
  • V. K. Khanna
  • G. K. Garg
Article

Abstract

Different cis acting elements of gamma kafirin gene from Sorghum bicolor var. M 35-1 were amplified and cloned using different combination of the primers. The amplified promoter was replaced with CaMV35S promoter of vector pCMBIA-1304 and resultant vector contained β-glucuronidase (gus) gene under the control of amplified γ-kafirin promoter. The resulting fusants were then transformed in to different explants of sorghum via particle bombardment. The regulation of uid gene expression was analyzed to find out the minimum required 5′ regulatory sequence and cis acting elements for the efficient expression. However no gus expression was detected in leaves of micropropagated plants, scutellum and calli at any stage of growth. The expression of gus, with pKaf gus-P4 gene construct, was detected in immature embryos and endosperm 20 days after pollination (DAP). The result suggest that at least three motifs (two GCN4 and one prolamin box) besides TATA and CATC boxes are required for the efficient expression of the kafirin gene of sorghum. The study shows that PCR based isolation of different motifs and regions can be used as an alternate to deletion analysis for observing the role of various motifs and their importance in the gene expression and regulation.

Keywords

gamma kafirin Gene regulation GLM Prolamin Promoter and transformation 

Abbreviations

DAA

Day after anthesis

DAP

Day after pollination

GLM

GCN4-like motif

Notes

Acknowledgement

The present investigation was a part of the project “Improvement of bread making quality and shelf life of grain sorghum” funded by NATP, Govt. of India. Financial assistance provided by NATP to AM, CSIR to AT and SB is duly acknowledged.

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Copyright information

© Springer Science+Business Media, Inc. 2007

Authors and Affiliations

  • Avinash Mishra
    • 1
    • 2
  • Akash Tomar
    • 2
  • Sangita Bansal
    • 2
  • V. K. Khanna
    • 3
  • G. K. Garg
    • 2
  1. 1.Molecular Biology and BiotechnologyCentral Salt and Marine Chemicals Research InstituteBhavnagarIndia
  2. 2.Department of Molecular Biology and Genetic EngineeringG. B. Pant University of Agriculture & TechnologyPantnagarIndia
  3. 3.Department of Genetics and Plant BreedingG. B. Pant University of Agriculture & TechnologyPantnagarIndia

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