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Cloning and Expression Analysis of Telomerase Reverse Transcriptase (AmTERT) Gene from Ammopiptanthus mongolicus
Sun Lichun1, Yang Ying1, Yu Tingqiao1, Sun Yuping1, Gao Haibo2, Wang Jinyu3, Men Jingyu1, Lu Cunfu1*, Chen Yuzhen1*
1Beijing Advanced Innovation Center for Tree Breeding by Molecular Design, College of Biological Sciences and Biotechnology, National Engineering Laboratory for Tree Breeding, Beijing Forestry University, Beijing 100083, China; 2College of Life Sciences, Linyi University, Linyi 276000, China; 3Analysis and Testing Center, Tsinghua University, Beijing 100084, China
Abstract: A segmented cloning method was used to clone a telomerase reverse transcriptase gene (AmTERT) of 3 459 bp in length from Ammopiptanthus mongolicus， and a systematic bioinformatic analysis of the telomerase reverse transcriptase gene (AmTERT) was carried out. The results showed that AmTERT encoded a protein of 1 153 amino acids with a relative molecular weight of 132.52 kDa and an isoelectric point of 9.49， which was a poorly stable hydrophilic protein. No transmembrane structure was found. The localization analysis results showed that it was probably located on the plasma membrane and in mitochondria， and also had NLS-monopartite and NLSbipartite nuclear localization signals. This amino acid sequence has a total of 147 potential phosphorylation sites and the result of amino acid alignment showed that the similarity between AmTERT and soybean TERT was as high as 75%. In addition， AmTERT protein has the conserved functional domains of telomerase reverse transcriptase (TRBD and RT)， and one putative Akt phosphorylation sites in AmTERT that conform to the consensus motif was predicted. Fluorescence quantitative PCR analysis showed that AmTERT expressed much more in roots than in stems and leaves. Furthermore， AmTERT expression increased under salt， drought， hot and cold stress， indicating that telomerase might be closely associated with genomic stability. Our research results implied that Ammopiptanthus mongolicus telomerase might have the non-telomere functions， and provided some new hints for further study to reveal the functions of plant telomerase.
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