中华蜜蜂工蜂cDNA文库的构建及ESTs测序分析
Construction of cDNA libraries and ESTs sequencing of Apis cerana cerana workers
张卫星** 郗学鹏 秦 明 王 帅刘春蕾 王红芳 胥保华***
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DOI:10.7679/j.issn.2095-1353.2016.075
作者单位:山东农业大学动物科技学院,泰安 271000
中文关键词:中华蜜蜂,cDNA文库,SMART技术,ESTs测序
英文关键词:Apis cerana cerana, cDNA libraries, SMART technique, ESTs sequencing
中文摘要:
【目的】 为了解中华蜜蜂Apis cerana cerana工蜂的抗逆性,构建了中华蜜蜂工蜂的cDNA文库,并对文库质量进行分析。【方法】 本研究利用SMART技术构建了中华蜜蜂工蜂的全长cDNA文库。【结果】 文库库容为3.6×106 cfu/mL,文库重组率为97%,插入片段长度多数分布在1 000 bp左右。挑取cDNA克隆进行EST测序,共进行了306个成功反应,软件拼接共得到234个单基因簇(Unigene),其中包括207个单拷贝(Singletons)序列及27个重叠群(Contigs)。使用Blastx将这些序列同GenBank等数据库进行查询、比对和注释,结果显示141条序列有相关同源性,其他序列没有明显的同源性,这也为我们发现新功能基因提供了可靠依据。【结论】 此文库的构建在中华蜜蜂功能基因的分离、克隆、筛选以及基因功能研究等方面具有重要作用。
英文摘要:
[Objectives] To build a cDNA library to improve understanding of how honey bee workers respond to adverse conditions and analyze the quality of the resultant library. [Methods] A cDNA library of Apis cerana cerana was constructed using the SMART technique. [Results] The library’s capacity was 3.6×106 cfu/mL, the recombination rate was 97% and the average length of inserts was approximately 1 000 bp. 306 ESTs were generated by ESTs sequencing. Additionally, 234 non-repetitive sequences were formed, including 207 singletons and 27 contigs after initial assembly. Using Blastx to query, compare and annotate these sequences with those in GenBank, revealed that 141 sequences could be assigned putative functions because they were homologous to known genes. Other sequences had no obvious homology, which suggests there is potential for the discovery of new functional genes. [Conclusion] The construction of a cDNA library has important benefits for cloning, screening and gene function research in Apis cerana cerana.