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胃癌相关基因GCRG123的克隆及其在大肠杆菌中的表达
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作者 伍银桥 王刚石 +2 位作者 王孟薇 吴本俨 尤纬缔 《世界华人消化杂志》 CAS 北大核心 2008年第35期3984-3987,共4页
目的:利用硫氧还蛋白融合表达系统表达胃癌相关基因GCRG123.方法:采用PCR技术从pGEM-T质粒上扩增出含完整ORF的GCRG123cDNA序列,将其克隆至硫氧还蛋白融合表达载体pET102/D-TOPO中,转化大肠杆菌BL21,经IPTG诱导表达重组融合蛋白,SDS-PAG... 目的:利用硫氧还蛋白融合表达系统表达胃癌相关基因GCRG123.方法:采用PCR技术从pGEM-T质粒上扩增出含完整ORF的GCRG123cDNA序列,将其克隆至硫氧还蛋白融合表达载体pET102/D-TOPO中,转化大肠杆菌BL21,经IPTG诱导表达重组融合蛋白,SDS-PAGE分析表达产物.结果:工程菌经IPTG诱导后,高效表达出相对分子质量约18700的重组融合蛋白.薄层凝胶扫描显示,其表达量占菌体总蛋白质的23.6%.结论:在大肠杆菌中成功表达了GCRG123重组融合蛋白. 展开更多
关键词 胃癌 基因 gcrg123 原核表达
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LINE-1 family member GCRG123 gene is up-regulated in human gastric signet-ring cell carcinoma 被引量:1
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作者 Gang-Shi Wang Meng-Wei Wang Ben-Yan Wu Xin-Yan Yang Wei-Hua Wang Wei-Di You 《World Journal of Gastroenterology》 SCIE CAS CSCD 2008年第5期758-763,共6页
AIM:To analyze the expression profiles of a human gastric-cancer-related gene, GCRG123, in human gastric signet-ring cell carcinoma tissues, and to perform bioinformatics analysis on GCRG123. METHODS:In situ hybridiza... AIM:To analyze the expression profiles of a human gastric-cancer-related gene, GCRG123, in human gastric signet-ring cell carcinoma tissues, and to perform bioinformatics analysis on GCRG123. METHODS:In situ hybridization was used to explore the GCRG123 expression pattern in paraffin-embedded gastric tissues, including 15 cases of signet-ring cell carcinoma, 15 of intestinal-type adenocarcinoma, and 15 of normal gastric mucosa. Northern blotting was used to analyze the differences in GCRG123 expression between stomach signet-ring cell carcinoma and intestinal-type adenocarcinoma tissues. Online software, including BLAST, Multalin and BLAT, were applied for bioinformatics analysis. National Center for Biotechnology Information (NCBI) and the University of California Santa Cruz (UCSC) databases were used for the analyses. RESULTS:The in situ hybridization signal appeared as blue precipitates restricted to the cytoplasm. Ten out of 15 cases of gastric signet ring cell carcinoma, normal gastric mucosal epithelium and pyloric glands showed high GCRG123 expression. Low GCRG123 expression was observed in gastric intestinal-type adenocarcinoma and normal gastric glands. Northern blotting revealed that GCRG123 was up-regulated in signet-ring cell carcinoma tissue but down-regulated in intestinal-type adenocarcinoma tissue. BLAST and Multalin analyses revealed that the GCRG123 sequence had 92% similarity with the ORF2 sequence of human long interspersed nuclear element retrotransposons (LINE-1, L1). BLAT analysis indicated that GCRG123 mapped to all chromosomes. GCRG123 was found to integrate in the intron-17 and-23 of Rb, 5' flanking region of IL-2 and clotting factor Ⅸ genes.CONCLUSION:GCRG123, an active member of the L1 family, was up-regulated in human gastric signet-ring cell carcinoma. 展开更多
关键词 LINE-1 胃癌 症状 癌细胞
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