Keywords

Wheat, High Affinity Potassium Transporter, Promoter

Abstract

Gene-specific primers were designed based on the common wheat HKT1 cDNA sequence (GenBank accession number: U16709), and a single clone BAC containing the HKT1 genome sequence was obtained by screening the Chinese spring BAC library. Sequencing primers were designed based on the known sequence of the 5′-end of HKT1, and the monoclonal BAC plasmid was used as a template for sequencing. The sequence was spliced by Sequencer software, and a 4192 bp sequence upstream of the start codon of the HKT1 gene was obtained. Neural Network Promoter Prediction (NNPP) software was used to analyze this sequence and predict the presence of 9 transcription start points. PLACE software analysis shows that this sequence has the basic elements of the promoter, TATA-box and CAAT-box, and contains multiple stress-inducible elements, such as salt-inducible elements GAAAAA, anti-freezing, water-deficiency, abscisic acid, cold-resistant elements CANNTG and CCGAC, etc. , injury-inducing element TGACY, tissue-specific expression elements ACTTTA and ATATT, etc. HKT1 promoter cloning showed that designing gene-specific primers to screen BAC libraries and obtaining gene promoter sequences through direct sequencing using monoclonal BAC plasmids as templates is a practical way to clone gene promoters from common wheat genomes.

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Original research done by Li Mengjun, Gao Xina, Fu Xiaoyi, Shi Zhanliang, Guo Jinkao

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