| 大鼠全长OBRb cDNA克隆及真核表达重组体的构建 |
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| 引用本文： | 王佑民,黎峰,程桦,傅祖植.大鼠全长OBRb cDNA克隆及真核表达重组体的构建[J].安徽医科大学学报,2002,37(5):333-335. |
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| 作者姓名： | 王佑民 黎峰 程桦 傅祖植 |
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| 作者单位： | 1. 安徽医科大学第一附属医院内分泌科,合肥,230022;2. 中山医科大学孙逸仙纪念医院内分泌科,广州,510120 |
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| 基金项目： | 广东省自然科学基金 (编号 :980 117)，卫生部基金 (标号 :98-1-0 91)资助课题 |
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| 摘 要： | 目的克隆SD大鼠全长长胞内段瘦素受体基因cDNA(OBRb cDNA)及构建其真核表达重组体,为进一步研究该基因打下基础.方法应用RT-PCR方法分段扩增全长OBRb cDNA,再将它们连接起来,并获得载有全长OBRb cDNA的阳性克隆质粒.将上述质粒中全长OBRb cDNA定向插入真核表达质粒pcDNA3中,筛选出阳性克隆.通过限制性内切酶酶切和核苷酸序列测定进行鉴定.结果重组的克隆质粒和真核表达质粒经限制性内切酶酶切后获得的片段大小均与理论值一致.结论成功地克隆了SD大鼠全长OBRb cDNA,并构建了真核表达重组体.
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| 关 键 词： | 克隆 分子 DNA 重组自由词 瘦素受体 |
| 文章编号： | 1000-1492(2002)05-0333-03 |
| 修稿时间： | 2002年3月29日 |
Cloning of complete OBRb cDNA from SD rat and constructing of its eukaryotic recombinant |
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| Wang Youmin,Li Feng,Cheng Hua et al.Cloning of complete OBRb cDNA from SD rat and constructing of its eukaryotic recombinant[J].Acta Universitis Medicinalis Anhui,2002,37(5):333-335. |
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| Authors: | Wang Youmin Li Feng Cheng Hua |
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| Abstract: | Objective To provide basis for investigating leptin receptor gene by cloning complete OBRb cDNA from SD rat and constructing its eukaryotic recombinant. Methods OBRb cDNA were amplified fragment by fragment using RT PCR and linked up into complete intact OBRb cDNA. And the positive clones with complete OBRb cDNA were screened out. Then complete OBRb cDNA was orientally inserted into pcDNA3 and its eukaryotic recombinant was obtained. At last, its eukaryotic recombinant was identified by digest of restrictive endonuclease and nucleotide sequencing. Results Sizes of fragments acquired were cosistant with theoretic values after cloned plasmid and eukaryotic recombinant were digested by endonuclease. Conclusion Complete OBRb cDNA was successfully cloned from SD rat and its eukaryotic recombinants were constructed. |
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| Keywords: | cloning molecular DNA recombinant |
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