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运用SMART技术成功构建了低温处理凡纳滨对虾仔虾的全长cDNA文库,文库的滴度为1.05×106pfu/mL,重组率94%,插入片段平均长度为1.0kb。随机选取116个克隆进行测序鉴定,获得111条有效序列,其unigene比例79.2%,涵盖了蛋白质合成、细胞骨架、核糖体结构、信号传导、代谢、细胞周期、能量产生与转换、转录、抗逆及防御、生物发育等10多种功能类别,其中与发育(10.34%)及抗逆及防御(10.34%)相关基因占20.68%。对测序获得的肌肉发育相关trioponinⅠ基因的3个不同拷贝进行了序列和进化比较分析。研究为发育和耐寒相关分子调控机制的探索创造了基础条件。
The full-length cDNA library of Litopenaeus vannamei was successfully constructed using SMART technology. The titer of the library was 1.05 × 106pfu / mL, the recombination rate was 94%, and the average length of insert was 1.0kb. One hundred and eleven valid sequences were randomly selected from 116 clones. The percentage of unigene was 79.2%, which included protein synthesis, cytoskeleton, ribosome structure, signal transduction, metabolism, cell cycle, energy production and transformation, transcription, resistance And defense and biological development. Among them, 20.68% were related to development (10.34%) and resistance and defense (10.34%). Three different copies of the trioponin I gene, which were related to muscle development, were sequenced and compared. The research has created the basic conditions for the exploration of developmental and cold-resistant molecular regulation mechanisms.