摘要
根据已知大鼠HPRT基因的外显子序列,从大鼠HPRT基因BAC中用酶切和PCR方法分别分离得到用于构建基因敲除载体的3.0kb和1.7kb的长臂(LongArm,LA)和短臂(ShortArm,SA),并克隆到pSL1180和pKO中.3.0kb长臂片段包含5′端部分序列和exon1部分序列,1.7kb短臂位于exon6和exon7之间.依次将neor、SA、LA克隆到pBS相应酶切位点,得到pBS-HPRT,再将LA-neor-SA片段由pBS中切出,克隆到pKO的KpnI和NotI位点之间,得到大鼠HPRT基因敲除载体pKO-HPRT.
Rat HPRT gene 3.0kb 5′flanking and part of exon1 DNA sequence which is separated by restriction enzyme digestion as Long Arm(LA), and 1.7kb exon6 to exon7 DNA sequence which is separated by PCR as Short Arm(SA),were used to construct HPRT gene knock out vector.Neor, SA,LA fragments cloned to suitable restriction enzyme sites of pBS respectively to construct pBSHPRT.Then the NeorSALA was cut from pBSHPRT and cloned to NotI and KpnI sites in pKO to construct pKOHPRT.The pKOHPRT was prepared to transfect rat fibroblasts and rat neural stem cells.
出处
《内蒙古大学学报（自然科学版）》
CAS
CSCD
北大核心
2002年第6期665-668,共4页
Journal of Inner Mongolia University：Natural Science Edition
基金
国家教育部重点项目(项目号:地方01019)
