Identification and Cloning of Proline Rich Region in Lentil( Lens Culinaris Medik.) With Ln-silico Analysis for Application in Genetic Engineering

Abstract

In present study a novel proline rich protein has been identified in lentil EST database using soyabean proline rich protein sequence as query sequence. Primer pairs were designed for this sequence and validated on selective Lens culinaris cultivars. The gene was 250bp in size and was cloned in ptz Vector for sequencing. tblastx result has shown 90 % similarity with chickpea EST sequence Accesion no. OR 398344 and 85% similarity with Lens culinaris EST sequence Accession no. OT 622346. Further Expression studies will help to study the role of Proline rich protein in response to various biotic and abiotic stress. Forward primer (PrpF) ACAAOCCACCTOTAOAOA and Reverse primer (PrpR) OTOT AAACTOOTOOCTOA were used in PCR amplification. The reaction was carried out at optimum condition required for amplification results in 250bp amplification and result was visualized by electrophoresis.Blast n results of the lentil sequence have shown 85 %similarity with Soyabean proline rich protein (NM001251664) and chickpea EST sequence (OR 398344). Pisum sativum (AJ233399), Gossypium hirusitum (AF044205) and Phaseolus vulgaris (AM158278) have also shown considerable similarity.