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| 体外转录法制备人野生型MDR1基因mRNA | |
| 引用本文： | 钱海鹏,向阳,田芳,李雷. 体外转录法制备人野生型MDR1基因mRNA[J]. 癌症进展, 2005, 3(3): 261-264 |
| 作者姓名： | 钱海鹏 向阳 田芳 李雷 |
| 作者单位： | 中国医学科学院,北京协和医院妇产科,北京,100730;中国军事医学科学院附属307医院,北京,100039;中国医学科学院阜外医院,北京,100037 |
| 摘 要： | 目的应用体外转录法大量制备人野生型多药耐药基因MDR1的mRNA,转染人造血干细胞后检测P糖蛋白表达.方法含人野生型MDR1基因的cDNA经酶切连接后构建可体外转录的模板,经体外转录得到mRNA,转染人造血干细胞,通过RT-PCR法检测mRNA及Western-bloting法检测P糖蛋白表达.结果转染后细胞mRNA相对含量(1.380)明显高于对照组(1.105)(P＜0.01);Western-bloting可见转染组MDR1基因编码的P-gp蛋白条带明显强于对照组.结论经体外转录法可大量制备人野生型MDR1基因的mRNA,并可成功应用于转染人造血干细胞,产生P糖蛋白表达. |
| 关 键 词： | 多药耐药基因 mRNA 体外转录 |
Construct a full-length mRNA for the human mutidrug resistance gene by method of transcribed in vitro |
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| Qian Haipeng,Xiang Yang,Tian fang,Li Lei. Construct a full-length mRNA for the human mutidrug resistance gene by method of transcribed in vitro[J]. Oncology Progress, 2005, 3(3): 261-264 | |
| Authors: | Qian Haipeng Xiang Yang Tian fang Li Lei |
| Abstract: | Obejective To construct a full-length mRNA for the human mutidrug resistance gene (MDR1)by method of transcribed in vitro,transfect it into human stem cell and test the expression of P-glycoprotein.Methods Digested plasmid with restriction enzymes to obtain the mdr1 cDNA fragment.Then,the fragment was inserted into clone vector and transcribed in vitro to gain the mRNA.Transfect the mRNA into human stem cell,test the expression by RT-PCR and Western-bloting.Results RT-PCR data showed mRNA was higher in transfected cells(1.380) than compare ones(1.105).Western-bloting showed P-gp was higher translated in transfected cells.Conclusion The full-length mRNA for the human MDR1 gene can be constructed by transcribed in vitro and used for transfected into human stem cell to express P-glycoproteins. |
| Keywords: | human mutidrug resistance gene mRNA transcribed in vitro |
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