| 镉诱导的蚯蚓全长cDNA文库的构建与初步鉴定 |
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| 引用本文： | 刘雪连,孙振钧,王冲,李帅章.镉诱导的蚯蚓全长cDNA文库的构建与初步鉴定[J].农业生物技术学报,2007,15(5):903-904. |
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| 作者姓名： | 刘雪连 孙振钧 王冲 李帅章 |
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| 作者单位： | 中国农业大学资源与环境学院,北京,100094 |
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| 基金项目： | 国家自然科学基金;教育部高等学校博士学科点专项科研基金;北京市生态学重点学科项目 |
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| 摘 要： | 采用人工土壤法，研究重金属镉(Cd2+)对成熟赤子爱胜蚓（Eisenia fetida）的急性毒性,表明蚯蚓对镉具有较强的耐受性。Trizol法提取蚯蚓RNA后，采用Clontech公司的CreatorTM SMARTTM cDNA文库构建试剂盒，成功构建了低剂量Cd2+（200 mg?kg-1）诱导的蚯蚓cDNA文库，其库容量为3.02×105 pfu/mL,扩增后滴度为8.67×109 pfu/mL，重组率为97.3％, 绝大多数的cDNA插入片段≥0?5 kb,平均长度≈1?0 kb。所够建的cDNA质粒文库容量及插入片段大小符合合格文库要求, 为进一步筛选蚯蚓体内解毒、免疫相关基因，寻找生物标志物奠定了基础。
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| 关 键 词： | 镉 耐受性 赤子爱胜蚓 cDNA文库 鉴定 |
| 文章编号： | 1006-1304（2007）05-0903-02 |
| 收稿时间： | 2006-11-23 |
| 修稿时间： | 2006-12-26 |
Construction and Primary Characterization of cDNA Library from Earthworm Induced by Cadmium |
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| LIU Xue-lian,SUN Zhen-jun,WANG Chong,LI Shuai-zhang.Construction and Primary Characterization of cDNA Library from Earthworm Induced by Cadmium[J].Journal of Agricultural Biotechnology,2007,15(5):903-904. |
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| Authors: | LIU Xue-lian SUN Zhen-jun WANG Chong LI Shuai-zhang |
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| Institution: | College of Resources and Environmental Sciences, China Agricultural University, Beijing 100094,China |
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| Abstract: | The acute toxicity of heavy metal Cd to earthworm (Eisenia fetida) was studied by the method of artificial soil test. It was shown that Eisenia fetida is tolerance specie which is applicable for accumulation indication. A cDNA library of Eisenia fetida was constructed .Total RNA was extracted from Eisenia fetida after induction with Cd2+ by Trizol regent. The full-long cDNA library was constructed successfully by using the SMART technique. The results showed that the primary titer of the constructed cDNA library was 3.02×105 pfu/mL, while that of the amplified library was 8.67×109 pfu/mL .The recombination rate was about 97.3%，and the length of most cDNA inserted in the library was about 300-3000bp.The library must be helpful to make clear the components of Eisenia fetida and establish the base for research into heavy metal detoxification and immune mechanisms at molecular level. |
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| Keywords: | cadmium tolerance Eisenia fetida cDNA library characterization |
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