Cloning and Analysis of Ice Recrystallization Inhibition Protein Gene TaIRI5 in Wheat
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    Abstract:

    In order to study the effect of ice recrystallization inhibition protein gene (TaIRI5) on wheat growth and development, we cloned the TaIRI5 gene from wheat cultivar‘Beijing841’by RTPCR technique. The gene was studied by bioinformatics analysis, tissuespecificity analysis, promoter activity analysis, subcellular localization analysis. The results showed that: (1) the TaIRI5 gene was successfully cloned, and the full length of TaIRI5 gene is 1 203 bp. The open reading frame was 858 bp, encoding 285 amino acids. The size of TaIRI5 protein is about 70.7 kD and the isoelectric point is 5.07. TaIRI5 is a hydrophobic protein. (2) Quantitative real time PCR analysis showed that TaIRI5 gene was expressed in roots, stems, leaves, stamens, pistils, glumes, seeds and the relative expression level of roots was the highest and stamens was the lowest. It is speculated that TaIRI5 gene plays an important role in the growth and development of wheat. (3) The promoter of TaIRI5 analysis showed that in addition to the core elements of CAATbox and TATAbox, the sequence also contains 9 light response elements and 6 hormone response elements as well as other elements. We have successfully constructed pCAMBIA1301 fusion expression vector containing GUS gene by three candidate promoters of TaIRI5 gene with different length of 498 bp, 999 bp and 1 500 bp. Tobacco transformation experiments showed that the three promoter candidates had strong activity with different expression patterns. (4) We have successfully constructed pCAMBIA1300TaIRI5GFP fusion expression vector containing GFP gene, subcellular localization analysis showed that TaIRI5 was located on the cell membrane. The results laid a foundation for the further study of TaIRI5 gene function in wheat.

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WANG Liuyi, ZHUANG Mengjia, YIN Jun, WANG Xiang. Cloning and Analysis of Ice Recrystallization Inhibition Protein Gene TaIRI5 in Wheat[J]. Acta Botanica Boreali-Occidentalia Sinica,2020,40(1):10-18

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  • Online: April 02,2020
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