| 摘要: |
| 在鳗弧菌侵染下, 利用SMART方法构建了青蛤的cDNA文库, 并采用高通量测序方法和BLASTX比对筛选出ESTs及免疫相关因子的基因。结果表明, 所构建的cDNA 文库的库容量为1.12×106, 插入片段均在500bp以上; 大于100bp的有效ESTs序列1113条, 平均长度725bp, 拼接后获得一致性序列420条, 其中包括126个叠联群和294个单拷贝EST, BLASTX比对后获得注释序列271条, 进一步筛选获得青蛤免疫相关因子基因序列45条, 为克隆其免疫防御相关因子的基因提供了重要的基础。 |
| 关键词: 青蛤, cDNA 文库, ESTs, 免疫相关因子基因 |
| DOI：10.11693/hyhz201301014014 |
| 分类号: |
| 基金项目:天津市科委应用基础与前沿技术重点项目资助, 12JCZDJC22800 号, 09JCZDJC19300 号 |
附件 |
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| CONSTRUCTION OF cDNA LIBRARY AND PRELIMINARY SCREENING OF IMMUNE-RELATED GENES IN CYCLINA SINENSIS |
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GAO Wei-Wei1, PAN Bao-Ping2
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1.Hexi District of Tianjin Community College For Staff;2.College of Life Science, Tianjin Key Laboratory of Animal and Plant Resistance, Tianjin Normal University
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| Abstract: |
| To investigate the molecular mechanism of immune of Cyclina sinensis, a cDNA library infected by Vibrio anguillarum was constructed based on SMART technique using Construction Kit (Clontech). The result of the test showed that the primary constructed cDNA library had a cloning capacity of 1.12×106 and the length of the insert fragments was above 500bp, therefore, the quality of the library could meet the requirements. By conducting high-throughput sequencing on white clones randomly selected from the library and informatics analysis of the sequences, 1113 expressed sequence tags (ESTs) out of which 126 contigs were obtained. Through the BLASTX on NCBI non-redundant protein database comparison, the total number of annotated sequences was 271 (64.5%), including 45 immune related sequences. These results could serve as the foundation of screening and cloning the full-length functional immune genes of C. sinensis. |
| Key words: Cyclina sinensis, cDNA library, Expressed sequence tags (ESTs), Immune related genes |
