| CELⅠ酶的粗提取及其活性检测 |
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| 引用本文： | 韩锁义,杨玛丽,盖钧镒,喻德跃. CELⅠ酶的粗提取及其活性检测[J]. 遗传, 2006, 28(9): 1112-1116 |
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| 作者姓名： | 韩锁义 杨玛丽 盖钧镒 喻德跃 |
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| 作者单位： | 南京农业大学大豆研究所/国家大豆改良中心/作物遗传与种质创新国家重点实验室, 南京 210095 |
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| 基金项目： | 国家重点基础研究发展计划(973计划)，中-法合作项目 |
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| 摘 要： | CEL I酶是第一个从真核生物中提取的用于高效特异切割DNA双链碱基错配和DNA扭曲的内切酶, 因而也是TILLING技术中用到的一种关键酶。文章对CELI酶的粗提取及其活性检测进行了研究。错配切割实验表明, CELI酶在包含有G→A点突变的杂合双链中, 能有效地在错配位点进行切割, 并可以通过ABI377测序仪获得直观的检测结果, 从而可以用于TILLING分析。
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| 关 键 词： | 点突变 活性 TILLING CELⅠ酶 |
| 文章编号： | 0253-9772(2006)09-1112-05 |
| 收稿时间： | 2005-09-26 |
| 修稿时间： | 2006-01-06 |
Crude Extraction and Activity Assay of CEL I |
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| HAN Suo-Yi,YANG Ma-Li,GAI Jun-Yi,YU De-Yue. Crude Extraction and Activity Assay of CEL I[J]. Hereditas, 2006, 28(9): 1112-1116 |
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| Authors: | HAN Suo-Yi YANG Ma-Li GAI Jun-Yi YU De-Yue |
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| Affiliation: | Soybean Research Institute, Nanjing Agricultural University; National Center for Soybean Improvement, Ministry of Agriculture; National Key Laboratory of Crop Genetics and Germplasm Enhancement, Nanjing 210095, China |
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| Abstract: | CEL I, extracted from celery, is the first known eukaryotic nuclease that cleaves DNA with high specificity at sites of base-substitution mismatch and DNA distortion. It is a key enzyme for TILLING research. Here we reported a crude extraction method and activity assay of CEL I. Incision at mismatches of single nucleotide suggested that CEL I can effectively detect DNA at G-->A base substitution and the result can be obtained from an ABI377 Sequencer. Therefore, the extracted enzyme can be used in TILLING. |
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| Keywords: | TILLING |
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