目的探讨载脂蛋白AⅠ-CⅢ-AⅣ(apolipoprotein AⅠ-CⅢ-AⅣ,Apo AⅠ-CⅢ-AⅣ)基因PstⅠ位点多态性与肝内胆管结石易感性(the happening of hepatolithiasis)的关系。方法应用PCR和限制性内切酶技术对64例肝内胆管结石患者和120例正常人A...目的探讨载脂蛋白AⅠ-CⅢ-AⅣ(apolipoprotein AⅠ-CⅢ-AⅣ,Apo AⅠ-CⅢ-AⅣ)基因PstⅠ位点多态性与肝内胆管结石易感性(the happening of hepatolithiasis)的关系。方法应用PCR和限制性内切酶技术对64例肝内胆管结石患者和120例正常人Apo AⅠ-CⅢ-AⅣ基因PstⅠ位点进行限制性片段长度多态性(restriction fragment length polymorphisms,RFLP)分析。结果肝内胆管结石组中A1A1、A1A2、A2A2基因型频率分别为0、4.7%、95.3%,正常对照组中A1A1、A1A2、A2A2基因型频率分别为0、10.0%、90.0%。肝内胆管结石组和正常对照组各基因型频率的比较(χ2=1.599,P=0.206)和等位基因频率的比较(χ2=1.531,P=0.216)差异均无统计学意义。结论 Apo AⅠ-CⅢ-AⅣ基因PstⅠ位点多态性可能不是肝内胆管结石的危险因素。展开更多
To identify, clone ,sequence and highly express the mature peptide gene of ApoA Ⅰ, total RNA was prepared from human fetal liver tissue. cDNA fragment encoding human ApoA Ⅰ was amplified by RT-PCR using specific pri...To identify, clone ,sequence and highly express the mature peptide gene of ApoA Ⅰ, total RNA was prepared from human fetal liver tissue. cDNA fragment encoding human ApoA Ⅰ was amplified by RT-PCR using specific primers, and then was inserted in pGEM-T vector. DNA sequencing indicates that the fragment is 729 base pairs in length and has 100% nucleotide homology with that of reported ApoA Ⅰ cDNA gene previously. The ApoA Ⅰ gene was cloned into pGEX 5X-1.The recombinant protein was expressed in E.coli DH5α, purified by glutathione-Sepharose 4B affinity chromatography and confirmed by SDS-PAGE. It was shown that the recombinant ApoA Ⅰ was expressed in E.coli, and the target protein amounted to 36% of total bacteria proteins. Cholesteryl ester transfer experiment showed that the recombinant ApoA Ⅰ was capable of promoting transfer of CE from HDL to LDL. Western blotting showed that the protein could react specifically with anti-ApoA Ⅰ antibodies.展开更多
文摘目的探讨载脂蛋白AⅠ-CⅢ-AⅣ(apolipoprotein AⅠ-CⅢ-AⅣ,Apo AⅠ-CⅢ-AⅣ)基因PstⅠ位点多态性与肝内胆管结石易感性(the happening of hepatolithiasis)的关系。方法应用PCR和限制性内切酶技术对64例肝内胆管结石患者和120例正常人Apo AⅠ-CⅢ-AⅣ基因PstⅠ位点进行限制性片段长度多态性(restriction fragment length polymorphisms,RFLP)分析。结果肝内胆管结石组中A1A1、A1A2、A2A2基因型频率分别为0、4.7%、95.3%,正常对照组中A1A1、A1A2、A2A2基因型频率分别为0、10.0%、90.0%。肝内胆管结石组和正常对照组各基因型频率的比较(χ2=1.599,P=0.206)和等位基因频率的比较(χ2=1.531,P=0.216)差异均无统计学意义。结论 Apo AⅠ-CⅢ-AⅣ基因PstⅠ位点多态性可能不是肝内胆管结石的危险因素。
文摘To identify, clone ,sequence and highly express the mature peptide gene of ApoA Ⅰ, total RNA was prepared from human fetal liver tissue. cDNA fragment encoding human ApoA Ⅰ was amplified by RT-PCR using specific primers, and then was inserted in pGEM-T vector. DNA sequencing indicates that the fragment is 729 base pairs in length and has 100% nucleotide homology with that of reported ApoA Ⅰ cDNA gene previously. The ApoA Ⅰ gene was cloned into pGEX 5X-1.The recombinant protein was expressed in E.coli DH5α, purified by glutathione-Sepharose 4B affinity chromatography and confirmed by SDS-PAGE. It was shown that the recombinant ApoA Ⅰ was expressed in E.coli, and the target protein amounted to 36% of total bacteria proteins. Cholesteryl ester transfer experiment showed that the recombinant ApoA Ⅰ was capable of promoting transfer of CE from HDL to LDL. Western blotting showed that the protein could react specifically with anti-ApoA Ⅰ antibodies.