【摘要】 采用原核表达载体pet-32-a+构建了羊驼皮肤组织cDNA文库。双链cDNA采用Universal Ribo Clonec DNA Synthesis System合成,并经过琼脂糖分级胶回收,去除了小于400 bp的片断;通过酶切载体的去磷酸化和插入片断的磷酸化,提高了重组率,蓝白斑筛选结果全部为阳性;库扩增后检测容量达2.25×107。随机挑取阳性克隆进行单引物测序分析,七个测序样品中获得两个与毛发生长相关的EST(expression sequence tag),分别为S100钙离子结合蛋白A3基因(S100 calcium binding protein A3,S100A3)的部分序列和高硫角蛋白关联蛋白基因(high sulfur keratin-associated protein,KRTAP3-3)的部分序列,说明所构建的文库具有较高的代表性。更多还原

【Abstract】 A cDNA library from the skin of alpaca in china was constructed using the prokaryotic expression vector pet-32-a+.All the cDNA fragments are normalized by 1 % agarose gel,and the fragments below 400 bp were discarded.The digested vector was processed by dephosphorylase and the cDNA fragments were processed by phosphorylase.The positive recombination rate was 100 % examined by blue/white colony.The library capacity was 2.25×107.Randomly selected 7 positive clones were sequenced by Shaihai Sangon bio-engineering company using T7 terminator primer.Two ESTs related to the wool growth were obtained,which are S100 calcium binding A3 gene and high sulfur keratin-associated protein gene.更多还原