CAJ | 학술논문

目的检测人肝癌细胞株 SMMC-7721中 POLD1基因 CDS 的突变情况,以期从 DNA 复制最关键酶的编码基因POLD1的突变为干预细胞恶性增殖提供新的思路.方法设计人 POLD1基因 CDS 特异性引物,以人肝癌细胞株 SMMC-7721总 RNA 为模板进行 RT-PCR 扩增,将扩增产物纯化后加入“腺嘌呤”并与 T 载体连接,将连接产物进行测序,与数据库中的序列进行比对.结果成功扩增了人肝癌细胞中 POLD1基因的 CDS 片段.SMMC-7721细胞中 POLD1基因的CDS 存在11个碱基替换,1个碱基缺失.结论人肝癌细胞 SMMC-7721中 POLD1基因的 CDS 发生了突变,进而导致其编码的氨基酸序列发生改变.
목적검측인간암세포주 SMMC-7721중 POLD1기인 CDS 적돌변정황,이기종 DNA 복제최관건매적편마기인POLD1적돌변위간예세포악성증식제공신적사로.방법설계인 POLD1기인 CDS 특이성인물,이인간암세포주 SMMC-7721총 RNA 위모판진행 RT-PCR 확증,장확증산물순화후가입“선표령”병여 T 재체련접,장련접산물진행측서,여수거고중적서렬진행비대.결과성공확증료인간암세포중 POLD1기인적 CDS 편단.SMMC-7721세포중 POLD1기인적CDS 존재11개감기체환,1개감기결실.결론인간암세포 SMMC-7721중 POLD1기인적 CDS 발생료돌변,진이도치기편마적안기산서렬발생개변.
@@@@Objective To analyze mutations in the protein-coding region of the POLD1 gene in the human hepatocarcinoma cell line SMMC-7721;this gene encodes an enzyme critical for DNA replication. Methods Total RNA was isolated from SMMC-7721 cultures,and the POLD1 gene coding region was amplified by RT-PCR,purified and A-tailed,and then cloned into a T-vector. Mutations were detected by sequencing. Results The protein-coding region of the POLD1 gene was successfully amplified and found to contain 12 mutations,11 of which were nucleotide changes and 1 was a nucleotide deletion. Conclusions SMMC-7721 en-codes a mutated POLD1 gene,and the mutations lead to changes in the protein sequence.