一个新的白血病相关基因LRP16全长cDNA的克隆、序列分析及表达特征  

Cloning and Expression Characterization of the Full Length cDNA for a Novel Leukemia-associated Gene LRP16

作　　者：韩为东[1] 于力[1] 楼方定[1] 王全顺[1] 赵瑜[1] 史子江[1] 焦宏远[2] 周建军[3] 

机构地区：[1]中国人民解放军总医院血液科,北京100853 [2]中国预防医学科学院病毒研究所,北京100052 [3]北京大学生命科学学院,北京100871

出　　处：《中国生物化学与分子生物学报》2001年第2期209-214,共6页Chinese Journal of Biochemistry and Molecular Biology

基　　金：国家自然科学基金!资助项目 (No.39970 82 )&&

摘　　要：克隆一个与白血病复发相关基因 (LRP1 6)的全长 c DNA序列 ,对其进行染色体定位、组织表达谱分析 ,并对该基因编码蛋白质进行原核表达 .首先用获得的一段 3kb DNA片段在 NCBI提供的 h ESTs数据库中进行电子杂交并对重叠克隆片段组装 ,再设计引物进行 c DNA末端快速扩增(RACE技术 ) .采用 Northern印迹方法进行组织表达分析 .以高通量基因组序列 (HTGS)数据库为基础进行染色体定位 .对构建的克隆菌用 IPTG诱导重组蛋白表达后进行 SDS- PAGE,同时对重组体测序确证 .钓取了该基因全长 c DNA、推导所编码的氨基酸序列 ,并将该基因定位于染色体1 1 q1 2 .2 .原核表达筛选获得了该基因重组子的一个缺失体 .对 LRP1 6基因全长 c DNA的序列分析提示 ,该基因可能编码两种 N端不同的蛋白质 ,且该基因的转录本可能存在一种丰度较低的剪接体 .To clone the full length cDNA of a novel leukemia relapse associated gene(LRP16),determine its chromosome location,analyse its tissue expression pattern and characterize the prokaryotic expression of this gene.Firstly,the human ESTs(Expression Sequence Tags)fragments obtained from electronic hybridization were assembled using a 3 kb DNA fragment cloned as probe,then designed the primers for rapid amplification of cDNA end(RACE).Tissue expression pattern were analysed by Northern blotting,and bioinformatic data of High Throughout Genomic Sequences(HTGS)were used for chromosome localization.Prokaryotic recombination protein was induced by using IPTG and bacterial protein electrophoresed on SDS PAGE.The inserted fragments of recombinant were confirmed by sequencing.The full length cDNA of the gene LRP16 was cloned and its translated amino acid sequence was deduced.This gene was localized on chromosome 11q12.2. One recombinant whose sequence had 30 bases deleted was obtained.LRP16 gene might produce two types of proteins and the only difference of the two was the length of their N terminus.This gene might have two types of splicing transcripts and the shorter lower transcripting.

关 键 词：白血病 相关基因 LRP16 重组蛋白 原核表达 CDNA 克隆 序列分析 

分 类 号：R733.702[医药卫生—肿瘤] R394.8[医药卫生—临床医学]