Rakesh Kumar Prajapat1*, Poonam Tiwari2, Puja Singh2 and Rekha Kansal2
1School of Agriculture, Suresh Gyan Vihar University, Jaipur
2ICAR-National Institute for Plant Biotechnology, Pusa, New Delhi
Email: rkp123000@gmail.com
Received-02.01.2022, Revised-19.01.2022, Accepted-27.01.2022
Abstract: Plants accumulate a set of defense proteins including lectins, proteinase inhibitors, amylase inhibitors etc. Lectins reversibly and non-enzymatically bind specific carbohydrates and this agglutination property makes them useful against various lepidopteran and homopteran insect pests. The isolated pigeonpea lectin (PPL) gene (~825 bp) was first cloned in pENTR-D-TOPO vector, subcloned into an expression vector (Gateway Destination vector pET300/NT-DEST) and transformed into BL21 DE3 pLysS competent cells of E. coli for protein expression studies. The PPL gene expression studies were carried out at different temperatures, IPTG concentrations and time intervals. The expression was maximum at 2.0 and 2.50 mM IPTG concentration at 37ºC for 5 hrs. The size of the protein was found to be around ~30 KDa. The expression was confirmed by SDS-PAGE and western blotting. Thus, transferring these defense genes under the control of tissue specific promoters will be an effective tool for sustainable insect pest management programme.
Keywords: Cloning, Expression, Insecticides, IPTG and Lectin
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