Plant stress tolerance related protein TaPK, and coding gene and application thereof
A gene and protein technology, which is applied to the plant stress tolerance-related protein TaPK and its encoding gene and application fields, can solve problems such as comprehensive improvement of stress tolerance of difficult plants.
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Embodiment 1
[0042] Embodiment 1, the cloning of TaPK
[0043] 1. Cloning of TaPK
[0044] Common wheat (Triticum aestivum L.) variety Triticum aestivum cv. Xiaobaimai grown in hydroponics for about 10 days can be obtained from the Institute of Crop Science, Chinese Academy of Agricultural Sciences, or from the National Germplasm Bank ( The serial number is ZM242); the literature referring to triticale is: Sun Haitao et al., Screening of interacting proteins of wheat TaDREB6 transcription factor, Chinese Agricultural Sciences, 2011, 44 (22): 4740-4747.; the literature mentioning triticale For: Isolation and molecular characterization of the Triticum aestivum L.ethylene-responsive factor 1 (TaERF 1) that increases multiple stress tolerance, Plant Mol Biol (2007) 65:719-732, Zhao Shi Xu, Lan Qin Xia, Ming Chen, Xian Guo Cheng, Rui Yue Zhang, Lian Cheng Li, Yun Xing Zhao, Yan Lu, Zhi Yong Ni, Li Liu, Zhi Gang Qiu, You Zhi Ma) The three-leaf stage seedlings were treated with NaCl for 2 hours,...
Embodiment 2
[0049] Embodiment 2, the impact of TaPK on plant stress tolerance
[0050] 1. Construction of recombinant expression vector
[0051] 1. Cloning of TaPK gene
[0052] A pair of primers (TaPK-121F and TaPK-121R) were designed according to the sequence of the TaPK gene, and SmaI and SpeI restriction sites were introduced at the ends of the primers respectively.
[0053] TaPK-121F: 5'-TTACCCGGGATGGACGAGGACGAGTACTCT-3' (SEQ ID NO: 3);
[0054] TaPK-121R: 5'-AGAACTAGTCGATCACAGCAGCTTCGG-3 (SEQ ID NO: 4).
[0055] The total RNA of white wheat leaves was extracted, cDNA obtained by reverse transcription was used as a template, and the above primers were used to amplify, and the obtained PCR product was subjected to 1.2% agarose gel electrophoresis, and its size was 1.8Kb. The PCR product is sent for sequencing, and the PCR product has the sequence 2 in the sequence listing, which is the TaPK gene.
[0056] Agarose Gel DNA Purification Kit Ver.2.0 (TaKaRa Company, Code No.: DV807A) ...
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