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目的:证实乙型肝炎病毒(hepatitis Bvirus,HBV)X基因一种新的变异方式.方法:从HBV慢性感染患者血清中提取HBVDNA,扩增X基因序列,克隆入pMD19T载体,选择阳性克隆进行DNA测序,与已知HBV基因相应序列比较该患者体内HBV基因变异位点以及变异形式.结果:从21例患者中共挑选74个克隆测序,测序结果提示54个克隆X基因下游大段缺失突变,长度达234nt,位于1601-1834nt处,另有1个克隆发生245nt缺失突变.发生缺失变异的病毒株同时存在G/A1515C、G1518C和A1585T替换突变,这两种突变具有联动特征.缺失突变株HBx仅编码76aa,其第44和45位编码为LL,具有特异性.结论:观察到一种X蛋白变异方式,这种大段缺失突变导致X蛋白下游编码序列丢失,其为X因子还是X蛋白以及这种变异是否为常态形式尚需进一步研究.
Objective: To confirm a new variation of X gene of hepatitis B virus (HBV) .Methods: HBVDNA was extracted from the serum of patients with chronic HBV infection and the sequence of X gene was amplified, cloned into pMD19T vector, and the positive clones were selected for DNA The HBV gene mutation sites and variants were compared with the corresponding HBV gene sequences.Results: A total of 74 clones were sequenced from 21 patients, and the sequencing results indicated that the downstream fragments of 54 clones had a large deletion deletion and the length Up to 234 nt at 1601-1834 nt and one clone at 245 nt deletion.The mutants of G / A1515C, G1518C and A1585T were also found in the strains with deletion mutation, and the two mutations were linked with each other.The deletion mutant HBx Encoding 76aa with codons 44 and 45 encoded as LL.Conclusion: An X protein variant was observed that led to the loss of the downstream coding sequence of the X protein, whether it is the X factor or the X protein and Whether this variation is the normal form needs further study.