糖尿病视网膜病变的二级康复干预:人血管生成素1的克隆和分析

Secondary rehabilitation preventive intervention for diabetic retinopathy: cloning and sequence analysis of human angiopoietin -1

目的:从人的胎盘中克隆血管生成素(angiopoietin-1,Ang-1)基因,并分析其结构特征。方法:提取人胎盘组织总RNA,用反转录聚合酶链式反应(RT-PCR)技术扩增出Ang-1基因片段,并将其克隆到PGEM-TEasy载体中进行序列分析。结果:获得高质量的胎盘组织总RNA。RT-PCR扩增出1.5kb的cDNA片段,成功构建了PGEM-T/Ang-1载体,Blast序列分析与GenBank中AY121504一致。结论:从人的胎盘中克隆Ang-1基因无突变,满足为进一步研构建pPIC9K/Ang-1毕氏酵母表达载体和表达蛋白质的需要。

AIM:To clone angiopoietin-1 (Ang-1)gene from human placenta,and analyze its structural characteristics. METHODS:Total RNA was isolated from human placenta,and then the Ang-1 cDNA fragment was amplified by reverse transcription-polymerase chain reaction(RT-PCR),and it was cloned into PGEM-T Easy vector for sequence analysis. RESULTS:High quality total RNA was isolated from human placenta. A 1.5 kb fragment of cDNA was amplified by RT-PCR,and PGEM-T/Ang-1 vector was successfully established. Blast sequence analysis was coincident with AY121504 in GenBank. CONCLUSION:Ang-1 cloned from human placenta has no mutation,which satisfies the needs for further study of establishing pPIC9K/Ang-1 Pichia Pastoris expression vector and expression protein.