Cloning and expression analysis of cysteine protease gene ( MwCP ) in Agropyron mongolicum

In this study, a cysteine protease gene (MwCP) from Agropyron mongolicum Keng was isolated using RACE. Sequence analysis indicated that MwCP was 1473 bp, and it contained a 1134-bp open reading frame, which encoded 377 amino acids with a 24-amino acid N-terminal signal peptide. The results indicated that the MwCP protein was a new member of the papain C1A family, and it was predicted to be an extracellular, secretory stable hydrophilic protein. The secondary structure of MwCP was mainly composed of α-helices and random coils, and the space structure primarily contained α-helices, β-sheets, and β-turns. Homology analyses showed the 98% homology between MwCP amino acids and a cysteine protease found in Triticum aestivum (GenBank accession No. AAW21813.1). Analysis of mRNA using semi-quantitative RT-PCR indicated that during a 48-h drought stress period, MwCP was expressed during the 4th hour, and the expression level peaked during the 6th hour before declining to 2 T.G.B.Y. Ao et al. ©FUNPEC-RP www.funpecrp.com.br Genetics and Molecular Research 15 (1): gmr.15017424 the original level. The results revealed that MwCP was involved in droughtresistant physiological processes of A. mongolicum. Moreover, the MwCP expression levels were highest in leaves, intermediate in roots, and lowest in stems.

[1]  Anil Kumar Singh,et al.  Cathepsin B cysteine protease gene is upregulated during leaf senescence and exhibits differential expression behavior in response to phytohormones in Picrorhiza kurrooa Royle ex Benth. , 2015 .

[2]  L. V. Savvateeva,et al.  Glutenase and collagenase activities of wheat cysteine protease Triticain-α: feasibility for enzymatic therapy assays. , 2015, The international journal of biochemistry & cell biology.

[3]  K. Yamaguchi-Shinozaki,et al.  Induced over-expression of AtDREB2A CA improves drought tolerance in sugarcane. , 2014, Plant science : an international journal of experimental plant biology.

[4]  R. Stolf-Moreira,et al.  Soybean physiology and gene expression during drought. , 2010, Genetics and molecular research : GMR.

[5]  X. Chang,et al.  Isolation and characterization of a gene encoding a polyethylene glycol-induced cysteine protease in common wheat , 2010, Journal of Biosciences.

[6]  N. Nassar,et al.  Drought tolerant stem anatomy characteristics in Manihot esculenta (Euphorbiaceae) and a wild relative. , 2010, Genetics and molecular research : GMR.

[7]  J. Tregear,et al.  Contrasting globulin and cysteine proteinase gene expression patterns reveal fundamental developmental differences between zygotic and somatic embryos of oil palm. , 2008, Tree physiology.

[8]  Tae-Geum Kim,et al.  Improvement of recombinant hGM-CSF production by suppression of cysteine proteinase gene expression using RNA interference in a transgenic rice culture , 2008, Plant Molecular Biology.

[9]  S. Arai,et al.  Gliadain, a gibberellin‐inducible cysteine proteinase occurring in germinating seeds of wheat, Triticum aestivum L., specifically digests gliadin and is regulated by intrinsic cystatins , 2007, The FEBS journal.

[10]  W. Hou,et al.  Molecular cloning and characterization of a granulin-containing cysteine protease SPCP3 from sweet potato (Ipomoea batatas) senescent leaves. , 2006, Journal of plant physiology.

[11]  Richard D Vierstra,et al.  The ubiquitin/26S proteasome pathway, the complex last chapter in the life of many plant proteins. , 2003, Trends in plant science.

[12]  P. Raymond,et al.  Changes in the Expression and the Enzymic Properties of the 20S Proteasome in Sugar-Starved Maize Roots. Evidence for an in Vivo Oxidation of the Proteasome1 , 2002, Plant Physiology.

[13]  H. Thomas,et al.  Sequencing, expression pattern and RFLP mapping of a senescence-enhanced cDNA from Zea mays with high homology to oryzain γ and aleurain , 1997, Plant Molecular Biology.

[14]  J. Mullet,et al.  A salt- and dehydration-inducible pea gene, Cyp15a, encodes a cell-wall protein with sequence similarity to cysteine proteases , 1995, Plant Molecular Biology.

[15]  K. Shinozaki,et al.  Arabidopsis DNA Encoding Two Desiccation-Responsive rd29 Genes , 1993, Plant physiology.

[16]  L. Qi.  Cloning and Expression Analysis of Cysteine Proteinase Genes(HbCP2 and HbCP3) from Para Rubber Tree(Hevea brasiliensis) , 2014 .

[17]  H. Rogers,et al.  Expression and localisation of a senescence-associated KDEL-cysteine protease from Lilium longiflorum tepals. , 2014, Plant science : an international journal of experimental plant biology.

[18]  He Hong-wu.  Cysteine Protease of Plant Potential Target of Herbicide , 2012 .

[19]  Wang Yujiong,et al.  Characterization, prokaryotic expression and purification of a rice cysteine protease OsCP2. , 2010 .

[20]  Z. Yun.  Full-length cDNA Cloning and Tissue Specified Expression Analysis of a Cysteine Protease in Brassica napus L. , 2008 .

[21]  Zhao Hong-fei,et al.  The Research on Mechanism of Programmed Cell Death , 2008 .

[22]  Yang Qing-chuan,et al.  Summary of cysteine protease in plants , 2005 .

[23]  S. Mohana,et al.  Degradation of the S. frugiperda peritrophic matrix by an inducible maize cysteine protease , 2005 .

[24]  B. Zagdańska,et al.  Multifunctional role of plant cysteine proteinases. , 2004, Acta biochimica Polonica.

[25]  Jiang Jian.  Cloning and Sequence Analysis of a Full-length Cysteine Proteinase cDNA in Gossypium hirsutum L. , 2004 .

[26]  Cui Ke.  Mechanisms of Programmed Cell Death and Its Relation to Development in Plants , 2000 .