其他题名Construction of a cDNA Library Using Gateway Technology and Analysis of Expressed Sequence Tags (ESTs) from Jerusalem Artichoke
杨少丽
; 秦松发表期刊生命科学研究
ISSN1007-7847
2018-10-31
卷号22期号:05页码:370-374+383
关键词菊芋cDNA文库Gateway技术表达序列标签(EST)基因注释Jerusalem artichokecDNA libraryGateway technologyexpressed sequence tag (EST)gene annotation
研究领域分子生物学
DOI10.16605/j.cnki.1007-7847.2018.05.004
产权排序(1)中国科学院烟台海岸带研究所; (2)
作者部门海岸带生物学与生物资源保护实验室
英文摘要A full-length cDNA library was constructed from the tuber of Jerusalem artichoke by recombinating the full-length cDNAs with the Gateway donor vector of pDONR222 for gene discovery and functional characterization. The cDNA library quality analysis showed that the capacity of original library was 5.76×106 CFU. The inserted fragments were mainly in the range of 1~3 000 bp and the recombination rate was 100% (24/24). These results indicated that the cDNA library had a high quality. EST sequencing was conducted and 2 639 high-quality ESTs were obtained, and 1 895 unigenes were obtained after assembly. NCBI database (NR) homologous alignment analysis showed that a total of 1 533 unigenes (80.9%) had significant homology with known genes. GO classification showed that the proportion of binding and catalytic activity were highest among the molecular function taxon. The cDNA library can be well used for functional genomics, novel gene screening, high-through EST sequencing and the preparation of cDNA microarray of Jerusalem artichoke.
中文摘要为丰富菊芋功能基因组学研究基础平台,以成熟期菊芋块茎为材料,将菊芋全长cDNA与Gateway供体载体pDONR222重组,构建了菊芋非剪切型全长cDNA文库。文库质量分析表明:未经扩增的原始文库库容量为5.76×10~6 CFU,插入片段大小主要为1～3000 bp,重组率为100%(24/24),达到了高质量文库的标准。利用该文库进行表达序列标签(expressed sequence tag, EST)测序,得到2639条高质量的EST序列,拼接后获得1895条非重复的唯一表达序列(unigene)。与NCBI的NR数据库同源比对分析表明,共有1533条unigene(80.9%)与已知基因有显著的同源性。GO分类结果显示:菊芋块茎表达基因在分子功能类群中,结合和催化活性所占比例最高。此cDNA文库将可用于菊芋功能基因组研究、新基因筛选、高通量EST测序以及菊芋cDNA芯片的制备等。
资助机构烟台市科技计划项目#2015ZH067; 国家自然科学基金项目#41001336
收录类别CSCD
语种中文
CSCD记录号CSCD:6357445
引用统计
文献类型期刊论文
条目标识符http://ir.yic.ac.cnhttp://ir.yic.ac.cn/handle/133337/24664
专题海岸带生物学与生物资源利用重点实验室_海岸带生物学与生物资源保护实验室
作者单位中国科学院烟台海岸带研究所;
推荐引用方式
GB/T 7714杨少丽,秦松. Gateway技术构建菊芋块茎全长cDNA文库及EST测序分析[J]. 生命科学研究,2018,22(05):370-374+383.
APA杨少丽,&秦松.(2018).Gateway技术构建菊芋块茎全长cDNA文库及EST测序分析.生命科学研究,22(05),370-374+383.
MLA杨少丽,et al."Gateway技术构建菊芋块茎全长cDNA文库及EST测序分析".生命科学研究 22.05(2018):370-374+383.
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