【摘要】 目的构建突触后致密物质-95(PSD-95)结构域PDZ1腺病毒重组体。方法以异硫酸氢胍-酚-氯仿一步法提取的总RNA为模板,采用反转录PCR(RT-PCR)法获得PDZ1的cDNA;与腺病毒穿梭载体用T4DNA连接酶连接;连接产物转化大肠杆菌JM109进行筛选、序列测定。结果①提取到未降解的总RNA;②经RT-PCR得到了PDZ1的cDNA;③酶切鉴定能观察到PDZ1和pAdTrack-CMV两条带;④PDZ1测序图谱与文献报道完全一致。结论获得了含目的基因PDZ1的腺病毒穿梭载体重组体。更多还原

【Abstract】 Objective To construct recombinant pAdTrack-CMV-PDZ1.?Methods Using the total RNA extracted from fresh rat hippocampus as template,the cDNA of PDZ1 was amplified by RT-PCR;the cDNA obtained was subsequently cloned into the shuttle vector pAdTrack-CMV(CLONTECH),and then,the sequence of the product was determined.?Results ①Total RNA was obtained.②The cDNA of PDZ1 was amplified by RT-PCR.③The recombinant was found containing two bands of pAdTrack-CMV/PDZ1 by restriction enzyme digestion.④The sequence of cloned PDZ1 was identical with that reported in GenBank.?Conclusion Recombinant of pAdTrack-CMV-PDZ1 is obtained.更多还原