Open AccessCloning and Expression Analysis of δ-OAT Gene from Saccharum spontaneum L
Author(s) -
Youli Yao,
Lei Xu,
Xiaowen Hu,
Yang Liu
Publication year - 2020
Publication title -
iop conference series. materials science and engineering
Language(s) - English
Resource type - Journals
eISSN - 1757-899X
pISSN - 1757-8981
DOI - 10.1088/1757-899x/780/3/032039
Subject(s) - gene , biology , genbank , genetics , accession number (library science) , coding region , open reading frame , cloning (programming) , gene expression , phylogenetic tree , sequence analysis , multiple sequence alignment , sequence alignment , peptide sequence , computer science , programming language
Full-length cDNAs of ornithines- δ -aminotransferase ( Ssδ-OAT ) were isolated from Saccharum spontaneum through homologous clone strategy. The sequence of this gene was then deposited in GenBank database with the accession number KX714115. Sequence analysis showed that the full length of the δ -OAT gene was 1373bp, the open reading frame was 1365 bp, it was inferred that coding 454 amino acids, the isoelectric point and the molecular weight of coded protein were 6.16 and 49.5 kD, respectively. According to the phylogenetic tree, the evolution of Ssδ-OAT corresponds with traditional biological classification. Its functional domain has a high conserved property in the process of biological evolution. Prokaryotic expression vector pEASY-E1-Ss δ -OAT was established and transformed BL21 (DE3) into E. coli after IPTG induction, showing a successful gene expression. The expression pattern analysis carried out by quantitative real-time PCR indicated that the gene expression level was significantly up-regulated under drought stress. The relative expression of the gene reached to the maximum under 6d treatment. It was the first time to obtain δ -OAT from Saccharum spontaneum L, Ssδ-OAT responded to water stress, it was inferred that δ -OAT played a role in resistance to osmotic stress.