人SAMHD1基因核心启动子区域的初步鉴定和分析
[Abstract]:Objective to identify the core promoter region of human infertility 伪 -motif domain and histidine / aspartic acid residue double conjoined domain (SAMHD1) gene, and to study the transcriptional regulation mechanism of SAMHD1. Methods Genomic DNA,PCR was extracted from Hep G2 cells to amplify the 937667553399 bp fragment upstream of the translation initiation site of SAMHD1 gene. The four amplified fragments were inserted into the p MD19-T vector. The DNA bands were identified by double enzyme digestion and then recovered into the p GL3-Basic vector. The results were confirmed by double enzyme digestion and DNA sequencing. The luciferase expression vector containing four fragments was cotransfected with pRL-TK into Hep G2 cells. The promoter activity of luciferase reporter gene was detected and analyzed. The transcription initiation site of SAMHD1 in Hep G2 cells was identified by 5'RACE. Results the genomic DNA.PCR was extracted from Hep G cells and then amplified. The results showed that 937667553399 bp fragment had been amplified. The results of double enzyme digestion electrophoresis showed that the four amplified fragments were successfully inserted into p MD19-T vector. Construction of luciferase expression vectors p GL3-937,p GL3-667,p GL3-553 and p GL3-399. by double enzyme digestion and DNA sequencing The detection of luciferase reporter gene activity showed that the core promoter of SAMHD1 was located in the 0- 399 region (the previous base of ATG was -1). The 5'RACE results showed that the transcription initiation site of SAMHD1 in Hep G2 cells was -101. Conclusion the core promoter of SAMHD1 is located in the region of -101 ~ 399 upstream of the translation initiation site, which lays a foundation for further study on the mechanism of transcriptional regulation of SAMHD1 in hepatocytes.
【作者单位】: 安徽医科大学免疫学教研室;安徽医科大学生物药物研究所;
【基金】:国家自然科学基金(编号:81372576) 安徽省自然科学基金(编号:1508085MH158) 安徽省博士后研究人员科研活动经费资助(编号:2015B066)
【分类号】:Q78
【相似文献】
相关期刊论文 前9条
1 高继平,郦永忠,,王宗阳,洪孟民;籼稻232蜡质基因转录起始位点的鉴定[J];遗传学报;1995年06期
2 杜耀华;王正志;倪青山;;基于滑动窗口的原核转录起始位点计算定位方法[J];生物物理学报;2006年05期
3 李志英;牟红珍;高丁梅;丁国平;马婷;王盛;;本氏烟Ⅰ型启动子的克隆及其转录起始位点分析[J];中国生物工程杂志;2014年01期
4 王智;转录起始位点的测定[J];基础医学与临床;1990年04期
5 李杰;张义全;高鹤;王丽;胡小许;周冬生;;利用优化的5′-RACE实验定位副溶血弧菌基因的转录起始位点[J];生物技术通讯;2014年03期
6 孙亮先;谢宪兵;桑庆亮;陈怀宇;陈朝阳;黄周英;;用5’LongSAGE标签分析蜜蜂Yps基因转录起始位点的多样性[J];泉州师范学院学报;2009年02期
7 庄海滨;朱景德;刘湘军;;人类全基因组范围的CpG岛的预测与分析[J];生物物理学报;2006年05期
8 孙亮先;黄周英;郑华军;游燕琳;;西方蜜蜂表皮蛋白基因apd-like转录起始位点的定位及cDNA序列的分析[J];昆虫学报;2011年02期
9 冯丽;任茂智;罗洪发;何光华;;分离植物目的基因全长cDNA和启动子的新方法——快速定位转录起始位点(RITIS)(英文)[J];分子植物育种;2006年01期
相关会议论文 前1条
1 孟繁疆;;从TSSP-TCM数据文件中提取转录起始位点[A];2007年中国农业工程学会学术年会论文摘要集[C];2007年
相关硕士学位论文 前3条
1 王丽雪;TNFα影响有机阴离子转运多肽1A2表达水平的分子机理研究[D];华南农业大学;2016年
2 侯婧逸;裂殖酵母全基因组范围内转录起始位点研究[D];上海交通大学;2012年
3 周艾彬;BMPRⅡ基因转录起始位点的确定及其近侧调控元件的鉴定[D];暨南大学;2007年
本文编号:2391305
本文链接:https://www.wllwen.com/kejilunwen/jiyingongcheng/2391305.html