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目的构建草珊瑚Sarcandra glabra叶片全长c DNA文库,进一步发掘与草珊瑚代谢及抗逆相关的基因,为草珊瑚功能基因的发现提供实验基础。方法以草珊瑚的嫩叶为实验材料,利用SMART法(即Clontech公司SMARTer试剂盒)构建草珊瑚全长c DNA文库。利用ABI3730 DNA序列分析测序获得大量的EST序列,利用生物信息分析方法对EST序列进行功能注释。结果构建了草珊瑚叶片的全长c DNA文库,经过鉴定草珊瑚c DNA文库的文库滴度为1.14×107 cfu/m L,平均插入片段为1 000 bp。利用该文库测序了221个单克隆,共获得EST序列177条,拼接出151个单一序列;利用NCBI数据库进行同源比对分析,共有119条(79%)与已知基因有显著的同源性,进行GO功能注释,显示其表达术语涉及了细胞生长,信号转导,蛋白质合成、转录,抗逆反应以及能量代谢等。结论构建了符合要求的草珊瑚叶片全长c DNA文库,并对相关EST序列进行了生物信息学分析,为草珊瑚基因组学研究提供一定的参考。
Objective To construct full-length c DNA library of Sarcandra glabra and further explore the genes related to metabolism and stress resistance of Sarcandra coraco, providing the experimental basis for the discovery of Sarcandra glabra functional genes. Methods Using the young leaves of Sarcandra glabra as experimental material, the full-length cDNA library of Sarcandra glabra was constructed by SMART method (ie, SMARTer kit from Clontech). A large number of EST sequences were obtained by ABI3730 DNA sequence analysis and functional annotation of EST sequences by bioinformatics analysis. Results The full-length c DNA library of Caragana was constructed. The titer of the cDNA library was 1.14 × 107 cfu / m L, and the average insert size was 1 000 bp. A total of 221 ESTs were sequenced using this library. A total of 177 ESTs were obtained and 151 single sequences were spliced. A total of 119 (79%) were homologous with known genes using NCBI database for homology analysis Sex, GO functional annotation, showing that the expression of terms related to cell growth, signal transduction, protein synthesis, transcription, anti-retrogradation and energy metabolism. Conclusion The full-length c DNA library of Caragana korshinskii was constructed and bioinformatics analysis of the related EST sequences provided some references for the research of genus Coralia.