膀胱癌组织某一特异性EST片段全长扩增及生物信息学分析

摘要

Objective To amplify the total length of a specific expressed sequence tag(EST)fragment of bladder cancer and to perform the bioinformatics analysis on it.Methods We collected 1 case of bladder cancer tissues which were pathologically confirmed bladder transitional cell carcinoma.The RNA from the bladder cancer tissues was extracted, and then we obtained the 5′end DNA sequence and 3' end DNA sequence of the EST fragment by RACE,and used the GeneTool to synthesize the results of DNA sequence.We analyzed the full length sequence of cDNA and its corresponding protein with bioinformatics,and submitted it to the NCBI to get GenBank accession number and protein ID by sequin soft -ware.Results We successfully got the 5′end and 3′end of the specific EST fragment,and used the GeneTool to synthe-size a full sequence of 1839bp,and it had integrated ORF,chromosome was located in 17p13.3.We got the GenBank ac-cession number,that was KR780251 in NCBI,and the ID of polypeptide sequence acquired protein translationed by ORF was AKO62675.Conclusion The gene of bladder cancer tissue-specific EST fragment is KR780251 with a full-length se-quence of 1839 bp and contains an intact ORF.The encoded protein AKO62675 may be a mutant from the normal bladder tissues.%目的 对膀胱癌组织某一特异性表达序列标签(EST)片段迸行全长扩增及生物信息学分析.方法 收集1例迸行手术的膀胱癌患者(病理证实为膀胱移行细胞癌)的癌组织,从新鲜的膀胱癌组织中提取RNA,利用RACE方法获取前期研究发现的某一特异性EST片段5′端和3′端DNA序列,通过GeneTool软件合成RACE测序结果.对全长序列及其相应蛋白迸行生物信息学分析,利用sequin软件编辑分析结果,并向NCBI提交获取基因号及蛋白号.结果 成功获得该特异性EST片段的5′端和3′端,利用GeneTool软件合成一个1839 bp的全长序列,初步分析其含有完整的开放阅读框(ORF),染色体定位于17p13.3.该序列在GeneBank获得基因号为KR780251, ORF翻译的多肽序列获得蛋白ID为AKO62675.结论 该膀胱癌组织特异性EST片段基因号为KR780251,全长序列为1839 bp,含有完整的ORF,其编码蛋白AKO62675可能是正常膀胱组织的基因突变体.