Cloning and characterization of lipoxygenase gene from germinating seedlings of green gram (Vigna radiata L.)

Autor: Dinakara Rao Ampasala, Kasi Viswanath Kotapati, Raveendra Anangi, Bhagath Kumar Palaka, Thyagaraju Kedam
Rok vydání: 2015
Předmět:
Zdroj: Indian Journal of Plant Physiology. 20:345-352
ISSN: 0974-0252
0019-5502
DOI: 10.1007/s40502-015-0186-z
Popis: Plant LOXs play a significant role in plant physiological processes such as growth, senescence, defence and stress-related responses. In the present study a full length lipoxygenase cDNA (VrLOX) has been cloned from green gram germinating seedlings using 5′- and 3′-RACE PCR method and nomenclatured VrLOX. The ORF (2601) encodes 867 amino acid residues with an approximate molecular weight of 97.43 kDa and an isoelectric point of 5.93. The VrLOX comprises of a putative PLAT domain and an iron-binding catalytic domain, and the sequence displayed a significant identity to known plant LOXs deposited in Protein Data Bank. Bioinformatics analysis demonstrated that this gene is most probably a LOX putative protein of type 13-LOX, and shared highest identity (91 %) with the Glycine max LOX-10. Enzymatic assay showed LOX activity was more on day 3 at pH 6.5 and higher MDA levels were observed on day 5 after germination. Semi-quantitative RT-PCR analysis indicated highest VrLOX mRNA expression on day 3 after germination, and could be induced by methyl jasmonate (MeJA). The 3-dimensional structure of the VrLOX was elucidated using in silico approach and molecular dynamics simulations.
Databáze: OpenAIRE
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