Cloning and Functional Analysis of Zinc Finger Protein Gene BrZFP1 in Brassica campestris
XU Jia1, ZHAO Yu-Hong2, HOU Xian-Fei3, TAO Xiao-Lei1, SUN Bai-Lin1, ZENG Rui1, ZHU Ming-Chuan1, SUN Wan-Cang1, WU Jun-Yan1, LIU Li-Jun1, LI Xue-Cai1,*, MA Li1,*
1 College of Agronomy/Gansu Provincial Key Laboratory of Aridland Crop Science/Gansu Key Lab of Crop Genetic and Germplasm Enhancement/Gansu Research Center of Rapeseed Engineering and Technology, Gansu Agricultural University, Lanzhou 730070, China; 2 Gansu Yasheng Agricultural Research Institute Co.Lt., Lanzhou 730070, China; 3 Institute of Economic Crops, Xinjiang Academy of Agricultural Sciences, Urumqi 830091, China
Abstract：Zinc finger protein transcription factors are one of the largest transcription factor families in plants, which participating in various biological processes such as apoptosis, autophagy and dryness maintenance, play an important role in regulating plant growth and development,and in response to adversity stress. In order to study the function of zinc finger protein gene in Brassica campestris, B. campestris variety 'Longyou 7' cDNA and gDNA were used as templates to clone the BrZFP1 gene (GenBank No. OK546027). Bioinformatics analysis of BrZFP1 gene sequence was carried out, and the transient transformation expression vector of Nicotiana tabacum was constructed for subcellular localization. Finally, the expression levels of BrZFP1 in leaves, growth cones and roots under low temperature, drought, high temperature and salt stress were detected by qPCR. The results showed that BrZFP1 encoded 160 amino acids, the predicted molecular weight of the protein was 17.094 kD, and the isoelectric point was 9.34. The gene had no intron and encoding a hydrophilic stable protein with a double conserved domain zf-C2H2_6. PlantCARE analysis results showed that the promoter region of BrZFP1 gene contained multiple cis-acting elements such as low temperature response, light response, meristem-related and hormone-related. The similarity between BrZFP1 amino acid sequence and homologous sequence of B. campestris was 98.75%. Subcellular localization showed that the protein was localized in the nucleus. Under low temperature stress, the change of BrZFP1 gene expression in the growth cone was consistent with the results of transcriptome sequencing, and the expression level showed an upward trend. Under different stress treatments, the expression of BrZFP1 gene in different parts increased to different degrees, drought and high temperature could strongly induce the expression of BrZFP1 gene. These results suggested that BrZFP1 gene might played an important regulatory role in abiotic stress in B. rapa. This study provides reference for further study on the function of BrZFP1 protein.
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