Construction of Expression Vector and Expression Analysis of Tobacco ERF Gene NtRAP2-7

doi:10.7668/hbnxb.2018.04.015

Abstract

Abstract: In order to study the function of tobacco NtRAP2-7 gene in abiotic stress response, NtRAP2-7 was cloned from K326 by homologous cloning. And the bioinformatics software was used to analyze the physicochemical properties, spatial structure and phylogenetic analysis of the encoded protein. Sequence analysis results showed that the gene contained 1 398 bp in length which encoded 465 amino acid residues and the predicted molecular weight of this protein was 51.16 ku. Based on 3-D modeling and secondary structure analysis, the protein consisted of three β-sheet and one α-helix and contained 58 phosphorylation sites. The subcellular localization prediction showed that the protein was mainly located in the nucleus and contained a monopartite nuclear location signal sequence. Phylogenetic analysis showed that NtRAP2-7 protein had the highest homology with Nicotiana sylvestris RAP2-7 protein sequence, which was 98%.The expression pattern analysis of the gene was carried out by qRT-PCR. Tissue expression analysis showed that the gene was expressed in the roots, stems, leaves and flowers, but had the highest expression level in roots. The expression levels of NtRAP2-7 was affected by abiotic stress treatments, indicating that the NtRAP2-7 was involved in the tobacco abiotic stress response. The pBI121-NtRAP2-7 overexpression vector was successfully constructed by double enzyme digestion, which laid the foundation for further study on the function of this gene in abiotic stress.

Key words: Tobacco, NtRAP2-7, Gene expression, Abiotic stress

CLC Number:

Q78
S572.03

CHEN Qian, ZHUO Wei, LUO Jing, YANG Shangyu, LU Liming, LI Liqin. Construction of Expression Vector and Expression Analysis of Tobacco ERF Gene NtRAP2-7[J]. Acta Agriculturae Boreali-Sinica, 2018, 33(4): 104-111. doi: 10.7668/hbnxb.2018.04.015.

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