摘要
目的 :本研究旨在鉴定质粒 pBlueBacHis2 -DPP上所携带的外源基因序列 ;建立带有人DPP基因的稳定克隆株 ;构建人DPP基因 5’端序列重组质粒 ,为表达纯化人DPP蛋白及其 5’端序列提供可参考的数据。方法 :用限制性内切酶分析并进行DNA序列测定质粒 pBlueBacHis2 -DPP上所携带的外源基因序列 ;以质粒pBlueBacHis2 -DPP为模板 ,构建人DPP基因 5’端序列重组质粒 ,并用蛋白序列分析软件包ANTHEPROT 4 5分析预测人DPP蛋白N端序列。结果和结论 :质粒pBlueBacHis2 -DPP上所携带的外源基因序列与基因文库上的人DPP基因序列相符合 ,建立带有人DPP基因的稳定克隆株pBV2 2 2 -DPP ;蛋白序列分析软件包分析预测人DPP蛋白N端序列氨基酸组成特点与人DPP全序列一致 ,体外即将表达的蛋白可能具有人DPP生物学功能的重组蛋白。
Objective: To test a recombinant plasmid (pBlueBacHis2-DPP) and to establish a store-stable cloning for human DPP gene and a 504-bp long fragment that covered 5’-terminus region of human dentin phosphoprotein gene (nt.1-504). Methods: Restriction enzyme assays in a recombinant plasmid (pBlueBacHis2-DPP) and sequence analysis from both two ends of inserting segment were analyzed.A 504-bp long fragment that covered 5’-terminus region of human dentin phosphoprotein gene (nt.1-504) ,amplified by PCR using pBlueBacHis2-DPP as the template ,was established .The general characteristics of the peptide encoded was predicted with molecular biology software ANTHEPROT 4.5. Results and Conclusion: Restriction enzyme assays revealed a roughly 2.3-kb long inserting segment in a recombinant plasmid (pBlueBacHis2-DPP) that was equal to the length of whole human DPP cDNA.Sequence analyses from both two ends of inserting segment showed that the tested nucleotides matched the human DPP 5’- and 3’-terminus sequences published on the Genebank.The recombinant plasmid pBV-222-DPP was generated in a store-stable cloning for human DPP gene.The structure of amino acids of the N-terminal Domain is the same as the DPP’s,which will lay a theoretical foundation for cloning and expressing the human DPP domain containing functional properties.
出处
《北京口腔医学》
CAS
2004年第4期198-201,共4页
Beijing Journal of Stomatology
