【摘要】 采用沙门氏菌以针刺法诱导家蝇三日龄幼虫,提取诱导后培养24 h的家蝇幼虫总RNA,进一步分离、纯化其mRNA,运用SMART技术构建家蝇幼虫cDNA文库。结果表明:原始文库的滴度为1.55×106pfu/mL,原始文库重组率为99.5%,文库扩增后滴度达1.27×1010pfu/mL。从扩增文库随机挑取10个噬菌斑克隆进行PCR扩增鉴定,结果显示所选的10个噬菌体克隆均含有重组的cDNA,插入片段大小为400~1 500 bp。更多还原

【Abstract】 The total RNA was extracted from Musca domestica larvae induced by Salmonella typhimurium.The mRNA was purified and the cDNA library of Musca domestica larvae was constructed by SMARTTM cDNA library Construction Kit(Clontech).The results showed that the primary titer of the constructed cDNA library was 1.55×106 pfu/mL,that of the amplified library was 1.27×1010 pfu/mL.The recombination rate was about 99.5 % and the length of most cDNA inserted in the library was 400—1 500 bp.更多还原