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| 应用基因组步移克隆小鼠Doc-1R基因组序列 | |
| 引用本文： | 生秀杰,姜莉,周伟强,王太一,张学. 应用基因组步移克隆小鼠Doc-1R基因组序列[J]. 中华医学遗传学杂志, 2001, 18(4): 314-316 |
| 作者姓名： | 生秀杰 姜莉 周伟强 王太一 张学 |
| 作者单位： | 1. 中国医科大学实验动物部;中国医科大学一院妇产科 2. 卫生部细胞生物学重点实验室 3. 中国医科大学一院妇产科 4. 中国医科大学实验动物部, |
| 基金项目： | 国家“九五攻关”项目(96-A230602)和国家自然科学基金(39980011) |
| 摘 要： | 目的：获得小鼠Doc-1R基因组序列。方法：根据小鼠Doc-1R基因的cDNA序列设计、合成特异引物，应用基因组步移策略对小鼠基因组步移文库进行扩增。以含有特殊接头（adaptor)的小鼠基因组步移文库作为模板，用巢式PCR法扩增目的片段。结果：应用此方法得到约1．5kb的目的片段，经测序分析证实Doc-1R基因克隆成功。此基因含有4个外显子、3个内含子，外显子与内含子接头符合GT－AG法则。结论：基因组步移方法简单、可靠、有效，是一种比较理想的克隆基因组片段的方法。 |
| 关 键 词： | 基因组步移 基因克隆 Doc－1R基因 基因组序列 |
| 修稿时间： | 2000-09-19 |
Cloning of the mouse Doc-1R gene by genomic walking |
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| X Sheng,L Jiang,W Zhou,T Wang,X Zhang. Cloning of the mouse Doc-1R gene by genomic walking[J]. Chinese journal of medical genetics, 2001, 18(4): 314-316 | |
| Authors: | X Sheng L Jiang W Zhou T Wang X Zhang |
| Affiliation: | Department of Laboratory Animal, China Medical University, Shenyang, Liaoning 110001 P.R.China. Xjsheng@hotmail.com |
| Abstract: | OBJECTIVE: To obtain the genomic sequences of the mouse Doc-1R gene. METHODS: Gene-specific primers were designed and synthesized based on the cDNA sequences of the mouse Doc-1R gene. With the use of genomic walking strategy, the mouse genomic walking library was amplified by the polymerase chain reaction(PCR). Mouse genomic library constructed with a special adaptor was utilized as a template to amplify the desired fragment by nested PCR. RESULTS: A desired fragment of 1.5 kb was obtained. Sequence analysis of the desired fragment confirmed that the genomic cloning of the Doc-1R gene was successful. This gene contains four exons and three introns. All of the splice donor/acceptor site sequences are in accordance with the consensus 'GT-AG' rule. CONCLUSION: The genomic walking strategy is simple, efficient and reliable; it is an ideal method of cloning genomic fragments. |
| Keywords: | genomic walking gene cloning Doc 1R gene |
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