摘要
为了克隆与白血病复发相关的新基因 (LRP15 )的全长cDNA序列 ,应用分子生物学及生物信息学技术对一段已知仅在白血病复发标本中被甲基化的 1.8kb长的DNA片段进行研究分析 ,通过美国生物信息中心 (NCBI)提供的hEST数据库进行电子杂交 ,并对获得的相互重叠的EST片段进行组装 ,设计引物进行cDNA末端快速扩增(RACE)。以高通量基因组序列 (HTGS)数据库及SAGE文库为基础 ,将获得的cDNA片段进行染色体定位及组织表达分析。结果表明 ,LRP15基因的cDNA序列全长 1718bp ,含 1个 780bp的开放读码框架 ,编码 2 5 9个氨基酸。该基因定位于染色体 3p2 4 ,在多种组织中表达。结论 :RACE技术是克隆新基因的有效方法 。
To clone the full length cDNA of a novel leukemia relapse associated candidate gene(LRP15), the human ESTs (Expression Sequence Tags) fragments obtained from electronic hybridization were assembled by a 1.8 kb DNA probe, which was only methylated in relapsed leukemia. Then the primers were designed for rapid amplification of cDNA end (RACE). Bioinformatic data of High Throughout Genomic Sequences (HTGS) and Serial Analysis of Gene Expression (SAGE) were used for chromosome localization and tissue expression analysis. The results showed that the full length cDNA of the novel gene had an open reading frame of 780 bp encoding a 259 amino acid protein of unkown functions. LRP15 gene expressed in many different tissues was localized in chromosome 3p24. It is concluded that RACE is an effective method to clone novel genes and LRP15 may be a leukemia relapse associated candidate gene playing an important role in carcinogenesis.
出处
《中国实验血液学杂志》
CAS
CSCD
2003年第1期22-26,共5页
Journal of Experimental Hematology
