Cloning and Characterization of a Heme Oxygenase-2 Gene from Alfalfa (Medicago sativa L.)

被引:6
作者
Fu, Guang-Qing [1 ,2 ]
Jin, Qi-Jiang [1 ,2 ]
Lin, Yu-Ting [1 ,2 ]
Feng, Jian-Fei [1 ,2 ]
Nie, Li [1 ,2 ]
Shen, Wen-Biao [1 ,2 ]
Zheng, Tian-Qing [3 ]
机构
[1] Nanjing Agr Univ, Coll Life Sci, Cooperat Demonstrat Lab Centrifuge Tech, Nanjing 210095, Peoples R China
[2] Nanjing Agr Univ, Beckman Coulter Ltd Co, Nanjing 210095, Peoples R China
[3] Chinese Acad Agr Sci, Inst Crop Sci, Natl Key Facil Crop Gene Resources & Genet Improv, Beijing 100081, Peoples R China
基金
中国国家自然科学基金;
关键词
Alfalfa; Heme oxygenase 2; Prokaryotic expression in E. coli; Gene expression; UP-REGULATION; OXIDATIVE STRESS; SOYBEAN PLANTS; NITRIC-OXIDE; EXPRESSION; ANTIOXIDANT; BILIRUBIN; LEAVES; ROOTS; METABOLISM;
D O I
10.1007/s12010-011-9343-7
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Heme oxygenase (HO, EC 1.14.99.3) catalyzes the oxidation of heme and performs vital roles in plant development and stress responses. Two HO isozymes exist in plants. Between these, HO-1 is an oxidative stress-response protein, and HO-2 usually exhibited constitutive expression. Although alfalfa HO-1 gene (MsHO1) has been investigated previously, HO2 is still poorly understood. In this study, we report the cloning and characterization of HO2 gene, MsHO2, from alfalfa (Medica sativa L.). The full-length cDNA of MsHO2 contains an ORF of 870 bp and encodes for 290 amino acid residues with a predicted molecular mass of 33.3 kDa. Similar to MsHO1, MsHO2 also appears to have an N-terminal transit peptide sequence for chloroplast import. Many conserved residues in plant HO were also conserved in MsHO2. However, unlike HO-1, the conserved histidine (His) required for heme-iron binding and HO activity was replaced by tyrosine (Tyr) in MsHO2. Further biochemical activity analysis of purified mature MsHO2 showed no HO activity, suggesting that MsHO2 may not be a true HO in nature. Semi-quantitative RT-PCR confirmed its maximum expression in the germinating seeds. Importantly, the expression levels of MsHO2 were up-regulated under sodium nitroprusside (SNP) and H(2)O(2) (especially) treatment, respectively.
引用
收藏
页码:1253 / 1263
页数:11
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