Cloning and Expression Analysis of Auxin Polar Transport PIN Gene 1 (CbPIN1) in Capsella bursa-pastoris

doi:10.7668/hbnxb.201750858

Abstract

Abstract: To probe the biological function of PIN1 gene in Capsella bursa-pastoris, the cDNA coding sequence of PIN1 (CbPIN1) gene was cloned by RT-PCR with the primers designed based on the reported Arabidopsis PIN1 amino acid sequence of Capsella bursa-pastoris in GenBank. The structure of CbPIN1 was analyzed via bioinformatics method. Also, CbPIN1 was expressed in prokaryotic cells. The CbPIN1 expression levels in Capsella bursa-pastoris tissues were detected using q-PCR method. In addition, the plant over-expression vector pBI121-CbPIN1 was constructed, and the transgenetic seedlings were also obtained. Sequencing analysis showed that the length of CbPIN1 gene cDNA was 1 869 bp, and the percentage of G+C was 49%. Further analysis indicated that CbPIN1 shared 93% homology with PIN1 of Arabidopsis. Information analysis demonstrated that the polypeptide encoded by CbPIN1 contained 622 amino acids. The weight and isoelectric point of CbPIN1 were 67.05 ku and 9.02, respectively. Moreover, CbPIN1 had 49 basic amino acids, 42 acidic amino acids, 241 hydrophobic amino acids and 157 polar amino acids. As a transmembrane protein, CbPIN1 contained 12 serine phosphorylation sites and one threonine phosphorylation site. The result of subcellular localization indicated that CbPIN1 was located in cytomembrane. q-PCR results suggested that CbPIN1 expressed in all tissues could be detected, and the highest expression site was root, and the lowest site was seed. The data indicated that CbPIN1 protein was 87.05 ku expressed in prokaryotic cells. In transgenetic seedlings of Capsella bursa-pastoris, CbPIN1 expression levels increased significantly. This research provided the foundation for further studying the function of CbPIN1 in Capsella bursa-pastoris organ development process.

Key words: Capsella bursa-pastoris, CbPIN1, Gene cloning, Expression analysis

CLC Number:

LIU Xiaozhu, LI Yinfeng. Cloning and Expression Analysis of Auxin Polar Transport PIN Gene 1 (CbPIN1) in Capsella bursa-pastoris[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2019, 34(1): 26-32. doi: 10.7668/hbnxb.201750858.

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