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1.55×10^6pfu/mL,原始文库重组率为99.5%,文库扩增后滴度达1.27×10^10pfu/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。
The total RNA was extracted from Musca domestica larvae induced by Salmonella typhimuritun. The mRNA was purified and the cDNA library of Musca domestica larvae was constructed by SMARTTM cDNA library Construction Kit （Clontech）. The results shawed that the primary titer of the constructed cDNA library was 1.55 × 10^6 pfu/mL, that of the amplified library was 1.27 × 10^10 pfu/mL. The recombination rate was about 99.5 % and the length of most eDNA inserted in the library was 400- 1500 bp.
出处
《吉林农业大学学报》
CAS
CSCD
北大核心
2009年第2期229-232,共4页
Journal of Jilin Agricultural University
基金
吉林省科技发展计划项目(20070578)
关键词
沙门氏菌
家蝇幼虫
CDNA文库
重组率
文库滴度
Salmonella typhimurium
Musca domestica larvae
eDNA library
recombination rate
titer of cDNA library
