| 两步扩增法在玉米抗大斑病相关基因片段分离中的应用 |
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| 引用本文： | 谷守芹,王海祥,范永山,韩建民,董金皋.两步扩增法在玉米抗大斑病相关基因片段分离中的应用[J].河北农业大学学报,2005,28(1):53-55,68. |
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| 作者姓名： | 谷守芹 王海祥 范永山 韩建民 董金皋 |
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| 作者单位： | 河北农业大学,真菌毒素实验室,河北,保定,071001;保定师范专科学校,河北,保定,071000;保定师范专科学校,河北,保定,071000;河北农业大学,真菌毒素实验室,河北,保定,071001 |
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| 基金项目： | 河北省自然基金项目(302333) |
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| 摘 要： | 对一步扩增法和两步扩增法在DDRT-PCR中的扩增效率进行了比较,结果发现,两种方法得到的片段大小都在150~1500 bp之间,但条带数量和清晰度不同。一步扩增法得到的扩增条带为30~45 条,条带较模糊,两步扩增法得到的扩增条带为40~60条,条带清晰。一步扩增法获得102 条玉米抗大斑病差异表达条带,只有55条带中的cDNA能够成功回收并产生有效扩增,差异条带回收率为53 9%,而两步扩增法获得了148条差异条带,其中138条带都能得到有效回收和扩增,回收率达93 2%。因此,两步扩增法更有利于差异条带的回收和二次扩增。
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| 关 键 词： | 玉米 两步扩增法 玉米大斑病 抗病相关基因 |
| 文章编号： | 1000-1573(2005)01-0053-03 |
The application of two- step amplification method in separation of expressed gene fragments in corn resistant to Setosphaeria turcica |
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| GU Shou-qin.The application of two- step amplification method in separation of expressed gene fragments in corn resistant to Setosphaeria turcica[J].Journal of Agricultural University of Hebei,2005,28(1):53-55,68. |
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| Authors: | GU Shou-qin |
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| Institution: | GU Shou-qin~ |
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| Abstract: | The amplification efficiency of one-step amplification method and the two-step amplification method in differential display reverse PCR (DDRT-PCR) were compared. It was found that the length of the amplified fragments by the two methods was all about 150～1000 bp,but the definition and the number of amplification band was different.30～45 bands were obtained by one-step amplification method, but the bands obtained by two-step were about 40～60.The bands obtained by two-step amplification method were much more legible than those obtained by one-step amplification method.102 differentially expressed bands related to the corn resistance to Setosphaeria turcica were acquired by one-step amplification method, in which 55 cDNA bands were successfully recovered, and the recovery percentage of the bands was about 53.9% while by two-step amplification method 148 amplification bands were acquired, in which 138 cDNA bands were successfully recovered, and the recovery percentage was 93.2%.In conclusion, the two-step amplification method was much more suitable for the mRNA differential display. |
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| Keywords: | corn two-step amplification method Northern corn leaf blight disease-resistance related gene |
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