长白山药用真菌鸡爪苓菌丝体全长cDNA文库的初步构建  

The Construction of the Full-length cDNA Library from Medicinal Fungi Chicken-claw-shaped Polyporus umbellate in Changbai Mountain

作　　者：廖鏖[1] 栾换东 李太元[1] 李艳茹[1] 许广波[1] 

机构地区：[1]延边大学农学院,吉林延吉133002

出　　处：《安徽农业科学》2014年第35期12418-12420,共3页Journal of Anhui Agricultural Sciences

基　　金：国家自然科学基金项目(31160014)

摘　　要：[目的]以长白山区鸡爪苓菌丝为材料,通过SMART技术构建全长cDNA文库.[方法]采用Trizol reagent试剂盒提取菌丝体总RNA,通过LD-PCR反转录合成双链cDNA,连接至载体λTriplEx2,构建鸡爪苓菌丝体全长cDNA文库.[结果]经测定,原始文库的滴度为1.66×106 pfu/ml,重组率为96.97％,扩增文库的滴度为1.504×1010pfu/ml,插入片段主要分布于500～2000 bp.[结论]成功构建了鸡爪苓菌丝cDNA,为鸡爪苓生长分化的分子机制研究提供理论基础.[Objective] With the mycelium of chicken-claw-shaped Polyporus umbellate in Changbai Moutain as material,a full-length cDNA library was constructed by using the SMART technology.[Method] Trizol reagent kit was used to extract the total RNA,which was reverse transcriptioned to synthetise the ds cDNA by LD-PCR.And then the ds cDNA was ligated to λTfiplEx2 to build the full-length cDNA library of Polyponus umbellatus mycelium.[Result] By the determination of experiment,the unamplified library titers was 1.66 × 106 pfu/ml,the recombinants rate was 96.97％.The titers of amplified library is 1.504 × 1010 pfu/ml,and the insert fragments were mainly distributed in 500-2 500 bp.[Conclusion] The full-length cDNA library of the Polyponus umbellatus mycelium was successfully built,which laid the foundation for the genetics and gene functional proteomics research of Polyponus umbellatus.

关 键 词：鸡爪苓 菌丝体 CDNA文库 构建 

分 类 号：S188[农业科学—农业基础科学]