MOLECULAR MARKERS OF HUMAN LIVER CANCER-NOVEL GENES DIFFERENTIALLY EXPRESSED

人类肝癌的分子标记——差异表达的新基因

• 批准号: 5201562
• 负责人: J LAUTENBERGER
• 金额: --
• 依托单位: DIVISION OF CANCER EPIDEMIOLOGY AND GENETICS
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/5201562
• 关键词: antibody; complementary DNA; computer assisted sequence analysis; endoplasmic reticulum; gene expression; genetic markers; histones; human genetic material tag; human tissue; liver neoplasms; membrane proteins; messenger RNA; molecular oncology; neoplasm /cancer genetics; northern blottings; nucleic acid repetitive sequence; nucleoproteins; polymerase chain reaction; protein sequence; secretory protein; tissue /cell culture; transcription factor

We have previously shown that rough endoplasmic reticulum (RER) from liver cell lines can give intact and enriched secretory protein mRNA (See project #Z01CP05739-01 LMO). Such mRNA was used as part of an assay system to determine which, if any, 3' cDNA fragments cloned by differential display correspond to secreted proteins. Out of 20 differentially expressed cDNA fragments, one was found novel by database sequence analysis and corresponding to a secretory protein by RER fraction-extracted RNA Northern analysis. We used 5' rapid amplification of cDNA ends (RACE) polymerase chain reaction (PCR) to clone the entire 3.7 kb mRNA from placenta mRNA, which was the only healthy tissue that expressed this novel clone. This gene contains a leader peptide sequence, a proline repeat, and seven potential membrane-spanning domains, according to computer analysis. The other 19 differentially expressed clones contain six complete novel genes, a histone-related gene, a transcriptional factor, and nuclear cytosolic proteins which are negative for RER localization, according to the previously discussed assay. We are currently focusing on our putative cell surface protein clone for protein expression, antibody production and patient sera/tissue screening to determine its diagnostic/ prognostic/therapeutic significance.

我们先前已经表明，粗面内质网（RER）从 肝细胞系可以产生完整和富集的分泌蛋白mRNA（参见 项目#Z01CP05739-01 LMO）。 这种mRNA被用作测定的一部分 系统，以确定哪些（如果有的话）3' cDNA片段被克隆 差异显示对应于分泌蛋白。 在20个差异表达的cDNA片段中，发现了一个新片段 通过数据库序列分析， 通过RER片段提取的RNA北方分析。 我们用5' rapid cDNA末端扩增（RACE）聚合酶链反应（PCR）， 从胎盘mRNA中克隆出完整的3.7kb mRNA，这是唯一的 表达这种新克隆的健康组织。这个基因含有一个 前导肽序列、脯氨酸重复序列和七个潜在的 跨膜结构域，根据计算机分析。 其他19 差异表达克隆包含6个完整的新基因， 组蛋白相关基因、转录因子和核胞质 蛋白质是负RER定位，根据 先前讨论的分析。 我们目前正专注于我们假定的 用于蛋白表达、抗体生产的细胞表面蛋白克隆 和患者血清/组织筛查，以确定其诊断/ 预后/治疗意义。