Construction of Expression Vector and Expression Analysis of Tobacco Protein Kinase Gene NtCIPK3
CHEN Qian1,2, YANG Shangyu1,2, ZHUO Wei1,2, LI Jiahao1,2, PENG Shuang1,2, WANG Jing1,2, LI Liqin1,2
1. College of Agronomy, Sichuan Agricultural University, Chengdu 611130, China;
2. National Demonstration Center for Experimental Crop Science Education, Chengdu 611130, China
Abstract：In order to study the function of CIPK3 in abiotic stress response,a CIPK family gene NtCIPK3 was cloned from common tobacco K326 by homologous cloning. The gene contained a 1 272 bp ORF encoding 423 amino acid. Protein sequence analysis indicated that the protein contained a transmembrane domain and belonged to a hydrophilic membrane protein. Protein alignment analysis revealed that NtCIPK3 contained a highly conserved N-terminal kinase region,a junction region,and a C-terminal regulatory region,with the highest homology to wild tobacco CIPK3,reaching 99%.Subcellular localization predictions showed that the protein was mainly located in the nucleus and had a bipartite typing nuclear localization signal sequence. The expression characteristics of this gene were analyzed by qRT-PCR. Analysis of tissue expression revealed that the gene was expressed in the roots,stems,leaves and flowers of tobacco and had tissue-specific expression and the expression level in leaves was significantly higher than that in other tissues,and it was speculated that it might function mainly in leaves.Meanwhile,NtCIPK3 gene was induced by low potassium,high salt,drought,ABA,H2O2,and low temperature treatment. It is speculated that NtCIPK3 gene played an important role in tobacco abiotic stress response. And successfully constructed the pBI121-NtCIPK3 overexpression vector,which laid the foundation for the future research of the gene under abiotic stress.
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2018, Vol. 33 
