| 人CD46基因启动子的克隆和启动特性研究 |
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| 引用本文： | 李国才,刘双喜,于峰,杜庆辉,王劲松,潘兴元,季明春,焦红梅.人CD46基因启动子的克隆和启动特性研究[J].扬州大学学报(农业与生命科学版),2012,33(1):10-13. |
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| 作者姓名： | 李国才 刘双喜 于峰 杜庆辉 王劲松 潘兴元 季明春 焦红梅 |
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| 作者单位： | 1. 扬州大学医学院,江苏扬州,225001
2. 江苏大学生命科学研究院,江苏镇江,212013 |
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| 基金项目： | 国家自然科学基金资助项目，江苏省自然科学基金资助项目 |
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| 摘 要： | 为研究人CD46(hCD46)基因启动子的启动特性,研制具有hCD46组织特异性表达特征的转基因小鼠,设计扩增hCD46基因启动子的PCR引物,用HeLa细胞基因组DNA为模板扩增hCD46基因启动子DNA片段,测序结果表明:其序列与GenBank中hCD46完整基因5′端某片段的同源性高达99.9%。用此DNA片段替换表达载体pcD-NA3EGFP中的CMV启动子,且在该片段与EGFP基因之间插入兔β-球蛋白基因的第2内含子RGI。重构的表达载体在2种鼠源细胞CHO和SP2/0中均可启动EGFP表达,表达量与人体内同源组织中hCD46的相似,说明该研究克隆了结构和功能正确的hCD46基因启动子。
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| 关 键 词： | 人CD46基因启动子 EGFP 启动特性 淋球菌 |
Cloning of human CD46 gene promoter and its promoting characteristic |
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| LI Guo-cai
,
LIU Shuang-xi
,
YU Feng
,
DU Qing-hui
,
WANG Jin-song
,
PAN Xing-yuan
,
JI Ming-chun
,
JIAO Hong-mei.Cloning of human CD46 gene promoter and its promoting characteristic[J].Journal of Yangzhou University:Agricultural and Life Science Edition,2012,33(1):10-13. |
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| Authors: | LI Guo-cai LIU Shuang-xi YU Feng DU Qing-hui WANG Jin-song PAN Xing-yuan JI Ming-chun JIAO Hong-mei |
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| Institution: | 1(1.Coll of Med,Yangzhou Univ,Yangzhou 225001,China; 2.Inst of Life Sci,Jiangsu Univ,Zhenjiang 212013,China) |
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| Abstract: | To study the promoting characteristic of human CD46 gene promoter,its DNA fragment was amplified by PCR using the template of the genomic DNA of HeLa cells.Sequence analysis revealed that the PCR product was 99.9% homologous to a DNA fragment in the 5′ end of hCD46 gene published in 2006.This PCR cloned hCD46 gene promoter was used to construct an eukaryotic expression reporting vector by using it to replace the CMV promoter in pcDNA3 EGFP.In order to enhance the expression level of interest genes,the 2nd intron of the gene encoding the rabbit β-globulin(RGI) was placed between the hCD46 gene promoter and the reporter gene of EGFP.The resultant vectors were transfected into 2 murine cell lines,CHO and SP2/0,respectively.FACS analysis demonstrated that the self-cloned hCD46 gene promoter could direct EGFP gene to express;the relative expression levels of EGFP in the 2 cell lines were similar to that of hCD46 in human homologous tissues.These data indicate that the self-cloned hCD46 gene promoter maintained its original promoting characteristic,thus could be used to establish hCD46 transgenic animals mimicking the expression properties of hCD46. |
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| Keywords: | human CD46 gene promoter EGFP promoting characteristic Syphilus |
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