Cloning and characterization of a plant defensin VaD1 from azuki bean
Cloning and characterization of a plant defensin VaD1 from azuki bean
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DOI：
10.1021/jf0402227
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发表时间：
2005-02-23
影响因子：
6.1
通讯作者：
Chen, CS
中科院分区：
文献类型：
--
作者：
Chen, GH;Hsu, MP;Chen, CS
重组绿豆防御素VrD1先前被证明具有抗真菌和抗野鼠的活性。为了研究VrD1的功能和调控，我们从豇豆VC6089A和红豆高雄6号中分离出植物防御素的基因组dna。对豇豆clefensin基因组DNA VaD1进行测序，并转化为VaD1 cDNA。从高雄6号角藤中纯化VaD1防御素，具有明显的同源性。对纯化后的VaD1进行了完整的氨基酸序列测定，发现与从VaD1 cDNA推导出的序列完全一致。VaD1是一种包含46个氨基酸和4个保守二硫键的碱性蛋白，与VrD1具有较高的序列同源性（78.3%）。VaD1抑制了尖孢镰刀菌、pisi尖孢镰刀菌、表皮葡萄球菌和鼠伤寒沙门氏菌的生长。VaD1对体外蛋白质合成和野鼠幼虫发育也有抑制作用，但活性低于重组VrD1。
A recombinant mungbean defensin VrD1 was previously shown to exhibit antifungal and bruchid-resistant activity. To study the function and regulation of VrD1, genomic DNAs of plant defensins were isolated from Vigna radiata VC6089A and azuki bean Vigna angularis Kao, Hsiung No. 6. The azuki bean clefensin genomic DNA VaD1 was sequenced and converted to VaD1 cDNA. VaD1 defensin was purified from Vigna angularis Kao Hsiung No. 6 to apparent homogeneity. The complete amino acid sequence of the purified VaD1 was determined and was found to be exactly the same as the sequence deduced from VaD1 cDNA. VaD1 is a basic protein containing 46 amino acids with four conserved disulfide bonds and shares high sequence homology (78.3%) with VrD1. VaD1 inhibited the growth of Fusarium oxysporum, Fusarium oxysporum f. sp. pisi, Staphylococcus epidermidis, and Salmonella typhimurium. VaD1 also inhibited in vitro protein synthesis and bruchid larval development, but was less active than the recombinant VrD1.