摘要
目的 对人纤溶酶原饼环区 - 5 (h PK- 5 )因子的 c DNA序列和氨基酸序列进行综合分析 ,为其基因工程表达载体的构建奠定理论基础。方法 从国际生化技术信息中心 (NCBI)的 Gene Bank数据库中调取人纤溶酶原的基因和全序氨基酸序列 ,结合文献报道结果获得 h PK- 5基因序列和氨基酸序列 ,利用蛋白质处理软件cd3d- win32、核酸分析软件 DNAclub和 SMS生物综合处理软件对 h PK- 5的基因序列和氨基酸序列进行综合分析。结果 选取 Eco RV(1890 )和 Sca I(10 91、1397)可以从纤溶酶原的基因序列中酶切获取一长为 4 93bp的基因片段 ,其中含有 h PK- 5序列 ;在 h PK- 5序列中间没有常见的限制性内切酶切割位点 ,酶切时可以保证外源基因序列的稳定性 ,h PK- 5基因序列中 ,G+C与 A +T相比略占优势 ,可以推断该基因序列的相对稳定性较高 ;氨基酸序列分析结果显示 ,在对 h PK- 5因子进行序列构建时 ,应根据其野生源蛋白的碱基及氨基酸序列特点 ,使用其随机频率的三联密码子 ,以保证后期基因表达蛋白具有高活性的可能性。结论 经多种生物软件分析 ,可以认为h PK- 5因子适合于利用基因工程技术体系制备 。
Objective To construct expression vector of human plasminogen kringle 5 (hPK-5) and to express the anti-angiogenesis in E.coli by analyzing the cDNA sequence and amino acid sequence of hPK-5.Methods The author fetched the full gene sequence and full amino acid sequence of plasminogen from web site of NCBI,combined with the results of literature about hPK-5,and analyzed above contents by using software of cd3d-win32,DNA club,etc.Results A 493 bp gene segment was obtained by digesting EcoRV(1890) and ScaI(1091,1397),including the gene sequence of hPK-5.There were no digest sites among hPK-5 gene sequence,and the amount of (G+C) was more than that of (A+T).These results concluded that the aim gene be stabilized.The amino acid analysis showed constructing the expression vector of hPK-5 must be based on its wild gene sequence′s character,using random codons to ensure the gene expression protein a high activity.Conclusion Lots of biological software analyses revealed that it is advisable to prepare hPK-5 factor by gene-engineering technology,thus,a relatively stable sequence can be obtained.
出处
《山西医药杂志》
CAS
2003年第5期421-424,共4页
Shanxi Medical Journal
基金
山西省归国留学人员基金资助项目 (2 0 0 0 - 4 1)
