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机构地区:[1]浙江医院消化内科,杭州310013
出 处:《胃肠病学和肝病学杂志》2005年第3期227-229,共3页Chinese Journal of Gastroenterology and Hepatology
基 金:国家科技部973计划项目G1999054105
摘 要:目的以乙型肝炎病毒(HBV)前C/C基因异质性来探讨在HepG2.2.15细胞体内是否存在HBV准种。方法依据Genebank中HBVayw亚型全基因序列,选择保守区域设计出特异性多聚酶链反应引物,自HepG2.2.15细胞培养上清中扩增HBV前C/C基因序列,克隆入pGEM-TEasy载体,挑选15个克隆测序以比较病毒的变异程度。结果测序结果发现HepG2.2.15细胞HBV前C/C基因存在异质性,碱基序列之间的同源性大于93%,突变类型全部为碱基替换。结论结果提示,长期培养HepG2.2.15细胞内有HBV准种共存,这种细胞模型内HBV异质性现象可能影响其在基因治疗等方面的应用。Objective To investigate the HBV quasispecees groups in the long-t erm cultured HepG2.2.15 cell. Methods A set of specific primers was synthesized according to the HBV DNA sequence of the subtype ayw in the GenBank, and the w hole pre-core/core region was amplified by the PCR method from the supernatant of HepG2.2.15 cell. Then the PCR products were subcloned into pGEM-T Easy vectors. Clones to be sequenced were selected, and then the sequence comparison was done to find the difference. Results One sequence of selected clones was found to be different; The homology of the nucleic acid sequence from the same patient was above 93.0%. The mutation type in this region was all substitution. Conc lusion There are HBV quasispecies groups in the HepG2.2.15 c ell. The HBV heterogeneity in the HepG2.2.15 cell may influence the application of the cell on hepatitis B gene therapy.
关 键 词:HEPG2.2.15细胞株 基因异质性 乙型肝炎病毒(HBV) HBV准种 多聚酶链反应 前C/C基因 全基因序列 培养上清 克隆测序 碱基序列 突变类型 长期培养 基因治疗 细胞模型 特异性 同源性 细胞内
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