关键词: 皮肤;β-防御素-3;逆转录聚合酶链反应

    摘 要:目的 克隆人β-防御素-3的基因. 方法 提取人皮肤组织的总RNA，反转录为cDNA作为模板，采用PCR的方法克隆人β-防御素-3基因.PCR产物连接入pGEM-T-EASY载体，转化DH-5α感受态细胞，蓝白筛选，对酶切及PCR鉴定含有目的片段的克隆进行测序. 结果 获得预期大小的PCR产物，经序列鉴定证实为人β-防御素-3基因. 结论 获得了人β-防御素-3基因.

    Cloning and sequencing of humanβ┐defensin┐3

    TUO Xiao-Ye，XU Ming-Da

    Department of Burn Surgery，Xijing Hospital，Fourth Military Medical University，Xi'an710033，China

    Keywords:skin;hBD-3;reverse transcriptase polymerase chain reaction

    Abstract:AIM To clone the gene encoding humanβ-de-fensin-3.METHODS Total RNA was extracted from hu-man foreskin tissue and complementary DNA was made by reverse transcription as template for polymerase chain reac-tion.PCR product of expected size was then inserted into pGEM-T-EASY cloning vector.Recombinant pGEM-hBD-3vector was transduced into competent cell DH-5α.Restriction analysis and PCR were performed to identify the clone con-taining the DNA fragment of interest，followed by sequencing with ABI CEI sequencer.RESULTS We obtained a259bp DNA fragment that is identical to the hBD-3cDNA sequence registered in GenBank.CONCLUSION hBD-3gene is suc-cessfully cloned from human foreskin tissue. ......