人血管生成素1真核表达载体的构建  

作　　者：唐莹[1] 刘学政[2] 李轶[2] 尚海峰[2] 王继红[3] 

机构地区：[1]辽宁中医药大学解剖教研室,辽宁沈阳110032 [2]辽宁医学院解剖教研室,辽宁锦州121001 [3]东北电力三公司职工医院眼科,辽宁锦州121001

出　　处：《中国医疗前沿（学术版）》2008年第5期34-35,共2页China Healthcare Innovation

摘　　要：目的构建人血管生成素1(angiogenin-1,ang-1)的真核表达载体。方法提取人总RNA,RT-PCR特异扩增人的Ang-1cDNA,与pGEM-T载体连接后转化大肠杆菌DH5α。筛选表达蓝白斑的阳性菌株扩增,提取质粒。选择正确的序列的产物,Xho I、BamH I双酶切产物和质粒,T4DNA连接酶连接,PCR扩增后转化大肠杆菌,抽提质粒进行测序。结果测序结果显示基因序列与人血管生成素1序列完全一致。结论成功构建了pEGFP/Ang-1真核表达载体。Objective To construct a kind of plasmid in which Ang-1 can express in eucaryotic cells. Methods Extract huamn total RNA, RT-PCR amplife the huamn Ang-1 cDNA, connect with pGEM-T vector and then convert E.coli DH5α. Choose the bacteria plaque witch express blue and white to amplife, then abstract the plasmid. Choose the plasmid witch espress the human Ang-1, use Xho I and BamH I to concis the PCR product and plasmid pEGFP, connect by T4DNA joining enzyme. Amplified by PCR and convert E.coli DH5, and abstrac...

关 键 词：人血管生成素1 克隆 真核表达载体 

分 类 号：R[医药卫生]