| 摘要: |
| 采用抑制性差减杂交方法进行了镉诱导下隆线溞生殖转化基因的抑制性差减cDNA文库的构建研究。结果显示, 其差异表达基因被富集了210倍左右, PCR 鉴定发现95%的克隆均含有200—800bp的插入片段; 测序获得的252个有效EST中, 与细胞分裂相关的有4个, 与细胞结构和功能相关的有52个, 与细胞代谢相关的有34个, 与信号传导相关的有16个, 与细胞防御相关的有10个, 与基因和蛋白质表达相关的有76个, 未知功能的EST有22个, 在GenBank中找不到任何同源序列的EST有38个。结果表明, 构建的差异表达cDNA文库具有很强的差减效率, 可较好地反映镉诱导下隆线溞差异表达的生殖转化基因信息。 |
| 关键词: 隆线溞, 镉, 生殖, 抑制性差减杂交 |
| DOI：10.11693/hyhz201204025025 |
| 分类号: |
| 基金项目:浙江省自然科学基金项目(Y3110038); 浙江省科技厅公益项目(2011C22086) |
附件 |
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| CONSTRUCTION AND ANALYSIS OF SUBTRACTED cDNA LIBRARY BY SUPPRESSION SUBTRACTIVE HYBRIDIZATION FROM THE REPRODUCTIVE CONVERSION GENES OF DAPHNIA CARINATA INDUCED BY CADMIUM |
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LUO Wen1, ZENG Wen-Tao1, WANG Dong-Fang2, ZHENG Da-Heng1, HAN Ji-Wei1
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1.School of Life Sciences, Shaoxing University;2.Xinchang County Laiyi Ecological Agriculture Development Co., Ltd.
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| Abstract: |
| Suppression subtractive hybridization (SSH) was used to construct the cDNA subtractive library from the reproductive conversion genes of Daphnia carinata induced by cadmium in this paper. The result showed that differentially expressed genes were enriched 210 times, 95% cloned sequences contained 200—800bp insertions in randomly selected positive clones, which were identified by PCR. 252 effective ESTs were obtained by sequencing, relevant to 4 belong to cell division genes, 52 belong to cellular structure and movement genes, 34 belong to metabolism genes, 16 belong to signal transduction genes, 10 belong to cell immune action genes, 76 belong to gene and protein expression associates genes and 22 belong to unknown functions; in addition, there are 38 no similar sequences in GenBank. The results suggested that the cDNA library had strong subtractive efficiency, SSH method was used to construct cDNA subtractive library could well reflect the differentiation gene information of the reproductive conversion of D. carinata induced by cadmium. |
| Key words: Daphnia carinata, Cadmium, Reproduction, Suppression subtractive hybridization |
