| 稻瘟菌诱导的稻叶脂氧合酶的纯化及部分氨基酸序列测定 |
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| 引用本文： | 侯占军,彭友良,Randeep Rakwal.稻瘟菌诱导的稻叶脂氧合酶的纯化及部分氨基酸序列测定[J].植物病理学报,1999,29(1):51-57. |
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| 作者姓名： | 侯占军 彭友良 Randeep Rakwal |
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| 作者单位： | 1 中国农业大学农业部分子植物病理学开放实验室, 北京 100094;2 Ibaraki University, Ibaraki 300-03 Japan |
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| 基金项目： | 美国洛克菲勒基金,霍英东优秀青年教师基金,国家教委《跨世纪优秀人才计划》基金,攀登计划植保项目,国家杰出青年科学基金,国家自然科学基金 |
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| 摘 要： | 作者对已报道的稻瘟菌诱导稻叶脂氧合酶CM-LOX 1和CM-LOX 2的纯化方法作了进一步改进和完善。改进后的纯化方法可缩短该脂氧合酶的纯化时间,并提高纯化倍数。利用改进的方法,作者从20 g非亲和性稻瘟菌小种接种稻叶中获得了电泳纯的酶蛋白CM-L OX 1 133 μ g和CM-LOX 2 208μg。在此基础上,采用特异性的蛋白酶endoproteinase Lys-C和溴化氰对该酶蛋白进行了裂解,回收其肽段,得到了CM-LOX 1的3个不同肽段共30个氨基酸和CM-L OX 2的7个不同肽段共87个氨基酸的序列,为最终克隆这2个酶的基因奠定了基础。
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| 关 键 词： | 稻瘟菌 脂氧合酶 纯化 氨基酸序列 |
PURIFICATION AND PARTIAL AMINO ACID SEQUENCING OF LIPOXYGENASES FROM RICE
INDUCED BY INCOMPATIBLE INFECTION OF Magnaporthe grisea |
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| Randeep Rakwal.PURIFICATION AND PARTIAL AMINO ACID SEQUENCING OF LIPOXYGENASES FROM RICE
INDUCED BY INCOMPATIBLE INFECTION OF Magnaporthe grisea[J].Acta Phytopathologica Sinica,1999,29(1):51-57. |
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| Authors: | Randeep Rakwal |
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| Institution: | 1 China Agricultural University, Beijing 100094;2 Ibaraki University, Ibaraki 300-03 Japan |
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| Abstract: | A previous procedure was modified to
shorten purification duration and enhance purification efficiency of CMLOX1 and CMLOX2 which
are two lipoxygenases induced in rice in response to incompatible infection of blast fungus.
Using this modified procedure, the authors obtained 133 g of CMLOX1 and 208 g of CMLOX2
with electrophoretic homogeneity from 20 grams of the pathogeninfected rice leaves.
Furthermore, the proteins were cleaved by LysCspecific proteinase and cyanogen bromide, and
partial amino acid sequences of the proteinderived peptides were determined. These involved
30 AA of 3 distinct peptides from CMLOX1 and 87 AA of 7 distinct peptides from CMLOX2. The
amino acid sequence information will greatly facilitate the cloning of cDNAs encoding the two
lipoxygenases. |
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| Keywords: | Magnaporthe grisea lipoxygenase purification amino acid sequence |
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