Scientia Agricultura Sinica ›› 2012, Vol. 45 ›› Issue (15): 3100-3107.doi: 10.3864/j.issn.0578-1752.2012.15.011

• HORTICULTURE • Previous Articles     Next Articles

Cloning and Expression of Cytosolic Glutamine Synthetase (GS1) in Cucumis sativus L. Under Low Nitrogen Conditions

 FENG  Zhuo, QIN  Zhi-Wei, WU  Tao, HE  Hong-Mei   

  1. 东北农业大学园艺学院，哈尔滨 150030
  • Received:2012-01-11 Online:2012-08-01 Published:2012-04-12

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Abstract: 【Objective】The purpose of this study is to clone full-length cDNA of key enzyme gene GS1 in plant nitrogen metabolism and to investigate its sequence characteristics and to analyze its expression under low nitrogen conditions.【Method】Sequences of primers were designed based on the Csa015274 gene coding region in the cucumber genome database using Primer Premier 5.0. The gene cDNA sequence was cloned from cucumber leaves using RT-PCR techniques; bioinformatics methods were used to analyze cDNA sequence obtained and putative amino acid sequence, qRT- PCR method were used to study the expression of GS1 gene under different nitrogen conditions. 【Result】 Cucumber GS1 gene (JQ277263) was got from cDNA by PCR. The gene is 1 071 bp, encoding 356 amino acids. Analysis of the nucleotide sequence by BLAST on line indicated that the homology of GS1 was up to 97% between Cucumis sativus L. and Cucumis melo L. The protein encoded by this gene is an unstable and hydrophobic protein with protein kinase C phosphorylation site, casein kinase II phosphorylation site, N-myristoylation site, tyrosine kinase phosphorylation site, and other active sites. Expression patterns analysis showed that the gene are down-regulated under low nitrogen conditions. And as the nitrogen concentration increased GS1 gene expression was increased. However, the expression of this gene is also inhibited at the high nitrogen level.【Conclusion】Gene GS1 was firstly isolated and characterized from cucumber. This gene had genetic characteristics similar with all species and it could provide reference for functional research.

Key words: Cucumis sativus L., GS1 gene, clone, sequence analysis, low nitrogen conditions

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