Lieto L D, Cothran E G
University of Kentucky, Dept. of Veterinary Science, Lexington 40546, USA.
Anim Biotechnol. 2001 May;12(1):87-97. doi: 10.1081/ABIO-100102981.
A cDNA library was built using RNA extracted from the skin tissue of an adult horse. The library was primed with oligo (dT) and sequences were directionally inserted in order to produce an expression library. The library has 5.8X 10(5) plaque forming units with 99.6% recombinant phage. The average insert size is 1.3 Kbp. Three hundred and thirteen expressed sequence tags (ESTs) were generated from sequencing of the 5 prime end of randomly selected skin cDNA clones. The ESTs were sequenced on an ABI 377 using Big-Dye chemistry. A similarity search was performed on each EST using the NCBI non-redundant protein database and 206 ESTs were putatively identified. Twenty six percent of the identified ESTs were redundant. The ESTs were categorized by function. The most frequently identified functional class was translational proteins.
使用从成年马皮肤组织中提取的RNA构建了一个cDNA文库。该文库用寡聚（dT）进行引物设计，并将序列定向插入以产生一个表达文库。该文库有5.8×10⁵个噬菌斑形成单位，重组噬菌体占99.6%。平均插入片段大小为1.3千碱基对。从随机选择的皮肤cDNA克隆的5'端测序产生了313个表达序列标签（EST）。使用Big-Dye化学法在ABI 377上对EST进行测序。使用NCBI非冗余蛋白质数据库对每个EST进行相似性搜索，推测鉴定出206个EST。鉴定出的EST中有26%是冗余的。EST按功能分类。最常鉴定出的功能类别是翻译蛋白。
