Malehorn D E, Borgmeyer J R, Smith C E, Shah D M
Agricultural Group, Monsanto Company, St. Louis, Missouri 63198.
Plant Physiol. 1994 Dec;106(4):1471-81. doi: 10.1104/pp.106.4.1471.
A cDNA clone encoding a basic thaumatin-like protein of Zea mays was recovered from a mid-development seed cDNA library. The gene, Zlp, encoded a protein that was nearly identical with maize zeamatin and alpha-amylase/trypsin inhibitor. Expression of Zlp mRNA was highest in the endosperm tissue of seed 4 weeks after pollination. Expression of zeamatin-like (ZLP) protein correlated with mRNA; also, a low basal level of ZLP expression in leaf was not appreciably induced by abiotic stresses. ZLP was expressed with its own signal peptide in insect cells and in transgenic Arabidopsis and tomato plants. ZLP was secreted in all three systems, with correct processing of the signal peptide. ZLP expressed in transgenic tomato was found to be partially subjected to a proteolytic cleavage after residue 180, by an unknown mechanism, to give a "nicked" isoform of ZLP. Purified ZLP from all three sources, as well as purified "nicked" ZLP from tomato, demonstrated fungal inhibition against Candida albicans and Trichoderma reesei, with marginal inhibition observed against Alternaria solani and Neurospora crassa.
从一个发育中期种子cDNA文库中获得了一个编码玉米碱性类甜蛋白的cDNA克隆。该基因Zlp编码的蛋白质与玉米甜蛋白和α-淀粉酶/胰蛋白酶抑制剂几乎相同。Zlp mRNA在授粉后4周的种子胚乳组织中表达最高。类甜蛋白(ZLP)的表达与mRNA相关;此外,叶片中ZLP的基础表达水平较低,非生物胁迫并未明显诱导其表达。ZLP在昆虫细胞以及转基因拟南芥和番茄植株中通过自身信号肽进行表达。ZLP在所有这三种系统中均有分泌,且信号肽加工正确。发现在转基因番茄中表达的ZLP在180位残基后通过未知机制部分受到蛋白水解切割,产生一种“带切口”的ZLP异构体。从所有这三种来源纯化得到的ZLP,以及从番茄中纯化得到的“带切口”ZLP,均表现出对白色念珠菌和里氏木霉的真菌抑制作用,对番茄链格孢和粗糙脉孢菌的抑制作用较弱。