Cloning and Expression Analysis of Lipid Transfer Protein LTP Gene in Carthamus tinctorius

doi:10.13560/j.cnki.biotech.bull.1985.2016.07.022

Abstract

Abstract: LTP（Lipid transfer protein）gene was cloned,its bioinformatics and expression were analyzed,providing the basis for the study the role of LTP gene in safflower resisting stresses. The sequence of LTP gene was cloned by RT-PCR technique,the protein characteristics were analyzed using bioinformatics,and the phylogenetic tree of LTP for safflower and other species was constructed. The expressions of LTP gene in the different tissues were analyzed using Real time-PCR. Sequence analysis showed that ORF of LTP was 294 bp,encoding a protein of 97 amino acids with a molecular weight of 7.46 kD,isoelectric point of 8.91. Safflower LTP protein contained a long signal peptide sequence of 29 amino acid residues,and the protein contained a serine phosphorylation site. Tertiary structure prediction indicated that the protein was a simple structure of 3 α-helix and 1 β-turn twining in random coils. Molecular evolution showed that the evolutionary relationship of LTP between Brassica of cruciferous and safflower was the most similar. Tissue-specific expression analysis of safflower LTP by fluorescence quantitative PCR showed that the gene expression levels in different tissues presented a significant difference,while high expression in seeds and flowers,and low expression in other tissues.

Key words: lipid transfer protein, safflower, Real-time PCR, bioinformatics analysis

HAN Yi-lai, CUI Qi, WU Yun-yun, GU Tian-yao, TENG Xiao-hua, HU Ren-ge, WU Guan-da, GUAN Li-li, LI Hai-yan, LI Xiao-kun. Cloning and Expression Analysis of Lipid Transfer Protein LTP Gene in Carthamus tinctorius[J]. Biotechnology Bulletin, 2016, 32(7): 146-151.

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