Molecular cloning and re-sequencing of trypsin inhibitor from a tropical timber tree red kelampayan

Neolamarckia mucropltylla (red kelainpayan) is a relatively unexplored fast-growing woody plant species with superior traits which can be used in reforestation programmes. Thus, the identification of the ability of it in resisting insect pests is needed. Insect pest resistant plants containing trypsin inhibitor, member of proteinase inhibitor can be useful in enhancing the economic traits of the plants. The presence of it helps in minimizing or excluding the usage of pesticides by utilizing the natural defence system of the plants. Therefore, the partial cDNA encoding for trypsin inhibitor from N. macrophylla was isolated and characterized by using in silico tools. Based on the nucleotide sequence analaysis, the cDNA isolated was miraculin-like gene which has the size of - 323 bp. The miraculin-like protein belongs to the Kunitz serine trypsin inhibitor family. It has the same function as trypsin inhibitor in protecting the plants from insect pests naturally. At the DNA level, the rapid and mass discovery of single nucleotide polymorphisms (SNPs) has becoming important in the studies of plant genetics. The results of re-sequencing indicate that there were 8 in silico SNPs identified from 7 samples of N. inacrophylla