【摘要】 NGAL(neutrophilgelatinase-associatedlipocalin)是lipocalin家族的一个新成员,可能是人类的一种新癌基因,但其在肿瘤中的表达调控机制尚不清楚。应用生物信息学手段对NGAL(X99133,包括外显子和内含子)进行分析发现,此区域内含有许多潜在的顺式作用元件,其中主要为增强子元件。对此设计了以下实验:应用PCR技术从食管癌细胞SHEEC基因组DNA中扩增不同长度的NGAL基因片段,将其连接到PGEMT-eacy载体中进行扩增,并测序。NCBI/BLAST分析确认扩增片段为NGAL基因所有,与实验所设计相吻合。在此基础上并应用KEGG/MOTIF检索分析系统对NGAL基因的上述区段进行了顺式作用元件定位分析,结果共鉴定出5个Score值=100分的顺式作用元件位点。这为NGAL基因增强子的鉴定提供了新线索。更多还原

【Abstract】 Neutrophil gelatinase-associated lipocalin(NGAL) is a novel member of the lipocalin family and may be a new human oncogene, but the regulation mechanism of NGAL was not clear in the cancer. Bioinformatics analysis shows that there are a lot of potential cis-acting elements,including some enhancers in the NGAL gene (X99133, including exon and intron). Therefore, the next tests were designed: 7 different length DNA fragments of the NGAL gene were amplified from genomic DNA of the esophageal cancer cell line SHEEC by PCR and inserted into pGEM-T easy vector and sequenced. NCBI/BLAST datbase analysis revealed that amplified fragments were the sequences of NGAL gene. Then cis-acting elements of the fragments were searched using KEGG/MOTIF system(http:∥www.genome.ad.jp ). 6 binding sites of 5 trans-acting factor(Sore=100) were identified. These results will offered an approach for further studying the enhancers of NGAL gene.更多还原