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PSD-95结构域PDZ1与腺病毒穿梭载体重组体的构建

Construction of recombinant pAdTrack-CMV-PDZ1
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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。 Objective To construct recombinant pAdTrack- CMV - PDZ1. Methods Using the total RNA extracted from fresh rat hippocampns 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 ① Total 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 report- ed in GenBank. Conclusion Recombinant of pAdTrack- CMV- PDZ1 is obtained.
作者 胡书群 纵艳艳 齐静 邰建敏 侯筱宇 张光毅
机构地区 徐州医学院生物化学与分子生物学研究中心
出处 《徐州医学院学报》 CAS 2005年第6期485-488,共4页 Acta Academiae Medicinae Xuzhou
基金 国家自然科学基金资助项目(30330190)
关键词 PDZ1 反转录PCR 腺病毒穿梭载体 序列测定 PDZ1 RT- PCR pAdTrack- CMV sequencing
分类号 Q78 [生物学—分子生物学]
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参考文献11

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徐州医学院学报
2005年 第6期

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