Expression Analysis and Functional Verification of TPS13 Gene in'Sensa-tion'Mango Fruit
In order to explore the function of terpene synthase gene in'Sensation'mango variety,In this study,'Sensation'mango pulp was used as the experimental material.A gene with the function of producing terpenes was cloned by homologous cloning technology,and the protein of the gene was induced and purified in the prokaryotic expression system.After adding the precursor substance for enzymatic reaction,the product of the gene was detect ed by gas chromatography-mass spectrometry(GC-MS).The results showed that a gene producing terpenes,named TPS13,could be amplified in the flesh of'Sensation'mango.The length of the gene was 1 692 bp,encoding 563 amino acids.TPS 13 was predicted to be hydrophilic and unstable protein by online website.The theoretical iso-electric point(pI)value was 5.51,indicating that the protein was acidic protein.By predicting its domain analysis TPS13 protein 52～540 amino acid interval contains an aspartic acid motif cd00684 domain,belonging to class Ⅰterpene cyclase superfamily.The identification of enzyme activity showed that the gene could not produce prod-ucts with substrates GPP,FPP and GGPP,and could catalyze the production of limonene and α-terpineol from monoterpene substrate(NPP).This experiment provided a reference for the study of mango terpenoids.
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