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牛MBL-A的血清水平检测及启动子区的甲基化鉴定
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作者 姜佳惠 李明涛 刘玉芬 《黑龙江畜牧兽医》 CAS 北大核心 2015年第11期95-97,100,289,共5页
为了检测牛血清中甘露聚糖结合凝集素(MBL)-A的浓度,并对其启动子区的甲基化情况进行分析,以分析MBL-A水平与牛的天然免疫力的相关性,试验采用酶联免疫吸附试验(ELISA)法对牛血清进行MBL含量的检测,以判定牛MBL-A的水平,同时对检测... 为了检测牛血清中甘露聚糖结合凝集素(MBL)-A的浓度,并对其启动子区的甲基化情况进行分析,以分析MBL-A水平与牛的天然免疫力的相关性,试验采用酶联免疫吸附试验(ELISA)法对牛血清进行MBL含量的检测,以判定牛MBL-A的水平,同时对检测合格的样品采集抗凝血进行DNA的提取,然后对启动子区进行克隆鉴定,再采用对甲基化敏感的内切酶,对启动子区的甲基化修饰位点进行初步判定,以期对牛MBL-A的遗传特征进行分析。结果表明:经ELISA检测,荷斯坦奶牛血清中MBL-A水平在12.30-109.69μg/L之间。PCR扩增得到牛MBL-A基因的启动子区序列,经酶切后得到2条片段,表明该酶切位点没有被甲基化修饰。 展开更多
关键词 甘露聚糖结合凝集素(MBL) 酶联免疫吸附试验(ELISA) 启动子 甲基化鉴定
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Identification of methylation profile of HOX genes in extrahepatic cholangiocarcinoma 被引量:7
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作者 Yi Shu Bing Wang Ji Wang Jian-Ming Wang Sheng-Quan Zou 《World Journal of Gastroenterology》 SCIE CAS CSCD 2011年第29期3407-3419,共13页
AIM:To identify methylation profile and novel tumor marker of extrahepatic cholangiocarcinoma(CCA)with high throughout microarray.METHODS:Differential methylation profile was compared between normal bile duct epitheli... AIM:To identify methylation profile and novel tumor marker of extrahepatic cholangiocarcinoma(CCA)with high throughout microarray.METHODS:Differential methylation profile was compared between normal bile duct epithelial cell lines and CCA cell lines by methyl-DNA immunoprecipitation (MeDIP)microarray.Bisulfite-polymerase chain reaction (BSP)was performed to identify the methylated allels of target genes.Expression of target genes was investigated before and after the treatment with DNA demethylating agent.Expression of candidate genes was also evaluated by immunofluorescence in 30 specimens of CCA tissues and 9 normal bile duct tissues.RESULTS:Methylation profile of CCA was identified with MeDIP microarray in the respects of different gene functions and signaling pathways.Interestingly,97 genes with hypermethylated CpG islands in the promoter region were homeobox genes.The top 5 hypermethylated homeobox genes validated by BSP were HOXA2(94.29%),HOXA5(95.38%),HOXA11 (91.67%),HOXB4(90.56%)and HOXD13(94.38%).Expression of these genes was reactivated with 5’-aza2’-deoxycytidine.Significant expression differences were found between normal bile duct and extrahepatic CCA tissues(66.67%-100%vs 3.33%-10%).CONCLUSION:HOXA2,HOXA5,HOXA11,HOXB4 and HOXD13 may work as differential epigenetic biomarkers between malignant and benign biliary tissues. 展开更多
关键词 DNA methylation EPIGENETIC Promoter microarray CHOLANGIOCARCINOMA
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