应用基因组步移克隆小鼠Doc-1R基因组序列  

Cloning of the mouse Doc-1R gene by genomic walking

作　　者：生秀杰[1,2] 姜莉[3] 周伟强[3] 王太一[1] 张学[3] 

机构地区：[1]中国医科大学实验动物部,沈阳110001 [2]中国医科大学一院妇产科 [3]卫生部细胞生物学重点实验室,沈阳110001

出　　处：《中华医学遗传学杂志》2001年第4期314-316,共3页Chinese Journal of Medical Genetics

基　　金：国家"九五攻关"项目 (96- A2 30 60 2 );国家自然科学基金 (39980 0 1 1 )&&

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 c DNA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 1.5 kb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 。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.

关 键 词：基因组步移 基因克隆 Doc-1R基因 基因组序列 

分 类 号：R392-33[医药卫生—免疫学]