Cloning and Sequence Analysis of the Amylase Gene from the Rice Pest Scirpophaga Incertulas Walker and its Inhibitor from Wheat (Variety MP Sehore)
Poonam Sharma, Pallavi R. Shankar, Geetha Subramaniam, Amit Kumar, Aanchal Tandon, C.G. Suresh, Meenakshi V. Rele and Lalitha Sunil Kumar
Division of Biochemical Sciences, National Chemical Laboratory, Pune 411008, India.
Abstract
Scirpophaga incertulas Walker (Lepidoptera: Pyralideae), commonly known as yellow stem borer, is a predominant monophagous pest of rice, which causes 5% to 30% loss of the rice crop. We report for the first time, the cloning and sequence analysis of the amylase gene of this pest. The cloned gene translates into a protein of 487 amino acids having a predicted molecular weight of 54,955 daltons and a theoretical pI of 5.9. The 3D structure of the amylase is predicted from its amino acid sequence by homology modeling using the structure of the amylase from Tenebrio molitor L (Coleoptera: Tenebrionidae). We also report the purification of a dimeric α-amylase inhibitor from a local variety of wheat MP Sehore that is specific for the amylase of this pest and does not inhibit human salivary amylase or porcine pancreatic amylase. The gene encoding this inhibitor has been cloned and its sequence has been analysed to find a possible explanation for this specificity.
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