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作 者:栾换东 蒋雨函 梁运江[1] 赵洪颜[1] 许广波[1]
出 处:《延边大学农学学报》2015年第4期275-280,共6页Agricultural Science Journal of Yanbian University
基 金:国家自然科学基金项目(31160014)
摘 要:以长白山鸡爪苓子实体为材料,采用Trizol改良法提取子实体总RNA,运用SMART技术构建鸡爪苓子实体全长cDNA文库。结果表明:原始文库的滴度为5.63×106 pfu/mL,重组率为98.73%;扩增后文库滴度为1.095×1010 pfu/mL,库容量约为2.19×1012。选取20个单克隆进行双向测序及分析,把成功获取的15条EST序列进行BLAST同源性比对分析,只找到了4条同源性序列,其余11条为鸡爪苓子实体新基因。With fruiting bodies of Chicken-claw-shaped Polyporus umbellatus from Changbai Moutains as materials,the total RNA was extracted using Trizol reagent kit and a full-length cDNA library was constructed by the SMART technology.The results showed that the unamplified library titer was 1.66×106 pfu/mL,the recombinants rate was 96.97 %.The titer of amplified library was 1.504×1010 pfu/mL.The library capacity was about 2.19×1012.The 20 monoclones selected were sequenced bi-directionally and analyzed,subsequently,obtained 15 high quality ESTs.Among the 15 ESTs,4 ESTs were homologoussequences to Nr database,the other 11 ESTs were novel sequences found from fruiting bodies of Chicken-claw-shaped Polyporus umbellatus from Changbai Mountains.
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