| 人野生型抑癌基因PTEN/MMAC1的巢式PCR法克隆、测序及表达载体的构建 |
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| 引用本文： | 田梅,刘林林,李修义.人野生型抑癌基因PTEN/MMAC1的巢式PCR法克隆、测序及表达载体的构建[J].吉林大学学报(医学版),2003,29(1):9-12. |
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| 作者姓名： | 田梅 刘林林 李修义 |
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| 作者单位： | 1. 吉林大学公共卫生学院 卫生部放射生物学重点实验室,吉林 长春 130021
2. 吉林大学第二医院放疗科 |
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| 基金项目： | 国家自然科学基金;39970229; |
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| 摘 要： | 目的 :克隆人野生型抑癌基因 PTEN/MMAC1 c DNA序列并构建其表达载体。方法 :利用RT- nested PCR法从正常人胎盘组织中扩增出约 1 2 0 0 bp的 DNA片段 ,与 p UCm- T载体连接 ,作全自动测序确证 ,并将其重组入 pc DNA 3.1载体中 ,构建为表达质粒 pc DNA- w P。结果 :对 PCR产物进行测序 ,序列基本正确。结论 :利用 RT- nested PCR法成功克隆了人野生型抑癌基因 PTEN/MMAC1c DNA序列并构建了表达质粒 pc DNA- w P。
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| 关 键 词： | PTEN/MMAC1 逆转录聚合酶链反应 巢式PCR法 表达载体 基因 抑制 肿瘤 基因表达 |
| 文章编号： | 1671-587X(2003)01-0009-04 |
Cloning and Sequencing of the Human Tumor Suppressor Gene PTEN/MMAC1 by RT-nested PCR and Construction of its Expression Vector |
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| Abstract: | Objective: To clone the cDNA of the human tumor suppressor gene PTEN/MMAC1 and construct its expression vector. Methods: About 1 200 bp DNA fragment was amplified from human placenta tissues by using RT nested PCR and was cloned into pUCm T vector after automatic sequenced, then the fragment was linked with expression vector pcDNA 3.1. Results:The PTEN/MMAC1 cDNA had been cloned correctly and its expression vector pcDNA wP also had been constructed. Conclusion: Human tumor suppressor gene-PTEN/MMAC1 cDNA had been cloned by RT nested PCR and its expression vector had been constructed successfully. The result makes the base for further study on the tumor suppression function of PTEN/MMAC1. |
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