Temporal and spatial expression analysis of gamma kafirin promoter from Sorghum (Sorghum bicolor L. moench) var. M 35-1
- 160 Downloads
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.
Keywordsgamma kafirin Gene regulation GLM Prolamin Promoter and transformation
Day after anthesis
Day after pollination
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.
- 9.Duffus CM, Cochrane MP (1992) Grain structure and composition. In: Shewry PR (ed) Barley: genetics, biochemistry, molecular biology and biotechnology. CAB International, Wallingford, pp 291–317Google Scholar
- 14.Shewry PR, Tatham AS, Halford NG (1999) The prolamins of the Triticeae. In: Shewry PR, Casey R (eds) Seed proteins. Kluwer Academic Publishers, Dordrecht, pp 35–78Google Scholar
- 18.Freitas F, Yunes JA, da Silvia MJ, Arruda P, Leite A (1994) Structural characterization and promoter activity analysis of gamma kafirin gene from sorghum. Mol Bio Mol Genet 245:177–186Google Scholar
- 21.Tomar A, Bansal B, Mishra A, Pandey D, Yadav MK, Khanna VK, Garg GK (2004) Enhancement of regeneration potential from immature embryo explants of grain sorghum (S. bicolor L.). In: Challenges & opportunity to harness the modern biology for socio-economic development in genomic era, symposium, G.B.P.U.A.T., Pantnagar, p 77Google Scholar
- 22.Vandana K (2002) Activity of amylase, protease and chitinase during early seedling growth in grain sorghum (Sorghum bicolor L.). MSc thesis, G.B.P.U.A.T., Pantnagar, IndiaGoogle Scholar
- 24.Ottoboni LMM, Leite A, Yunes JA, Targon MLPN, Souza FGA, Arruda P (1993) Sequence analysis of 22 kDa like α-coixin genes and their composition with homologous zein and kafirin genes reveals highly conserved pritein structure and regulatory elements. Plant Mol Biol Int J Mol Biol Biochem Genet Eng 21:765–778Google Scholar
- 25.Mazhar H, Chandrashekar A (1993) Differences in kafirin composition during endosperm development and germination in Sorghum cultivar of varying hardness. Cereal Chem 70:667–671Google Scholar
- 28.Thompson WF, Murray MG (1981) Protein and nucleic acids. In: Stumpf PK, Cohn EE (eds) The biochemistry of plants – a comprehensive treatise, 6 Academic Press, New York, USA, p 1Google Scholar