论文部分内容阅读
目的克隆Epstein-Barr(EB)病毒A73基因,并分析其编码区序列变异情况,为进一步研究其生物学功能提供基础。方法从7例鼻咽癌组织中分别提取其总RNA,以RT-PCR扩增A73基因编码序列,将其克隆入pGEM-T-Easy载体进行测序,以Blast软件在Genbank数据库中对其进行序列分析。结果成功克隆了高发区鼻咽癌的A73基因,序列分析表明该基因编码序列第45nt发生A→C的碱基替换,该变异位于第VB外显子的157154nt,未引起相应第15位氨基酸的替换(CCA→CCC,P→P),其变异频率为7/7。结论高发区鼻咽癌EB病毒A73基因的编码序列存在变异,该变异属于鼻咽癌相关的多态性,有可能参与EB病毒某亚型的构成。
Objective To clone the A73 gene of Epstein-Barr (EB) virus and analyze the variations of its coding region sequence, so as to provide a basis for further study on its biological functions. Methods The total RNA was extracted from seven NPC tissues. The coding sequence of A73 gene was amplified by RT-PCR, cloned into pGEM-T-Easy vector and sequenced by Blast software in Genbank database analysis. Results The A73 gene was successfully cloned from nasopharyngeal carcinoma in high incidence area. Sequence analysis showed that A → C base substitution occurred at the 45th of the coding sequence. This mutation was located in 157154nt of exon VB and did not cause the corresponding 15th amino acid Substitution (CCA → CCC, P → P), the frequency of variation is 7/7. Conclusion The coding sequence of Epstein-Barr virus A73 gene in nasopharyngeal carcinoma in high-incidence areas is variable. This variation belongs to the polymorphism of nasopharyngeal carcinoma and may be involved in the formation of a subtype of Epstein-Barr virus.