假单胞菌亮氨酸氨肽酶基因克隆及生物信息学分析

Cloning and Bioinformatics Analysis of LAP Gene from Pseudomonas

为探索假单胞菌亮氨酸氨肽酶(leucine aminopeptidase,LAP)编码基因LAP的结构和功能特征及其在生长发育中的作用,克隆假单胞菌Cr13菌株的LAP基因,并对其进行生物信息学分析。首先,克隆假单胞菌亮氨酸氨肽酶的一个DNA片段;其次,以此为基础,通过2次染色体步移(genome walking)对假单胞菌LAP基因全长序列进行克隆;最后,采用生物信息学软件对LAP进行分析。结果表明:Cr13 LAP基因全长为1491 bp。推测LAP基因定位于线粒体,具有保守的细胞质氨肽酶模体NTDAEGRL,在其启动子区域(426 bp)发现了基本元件TATA-Box和CAAT-box及一系列潜在的顺式作用元件。LAP全长序列的克隆为将来深入研究该基因的功能奠定了基础,LAP启动子序列的克隆与分析为进一步研究假单胞菌Cr13 LAP的表达调控提供了数据依据。

To explore the molecular structure and functional characteristics of LAP gene encoding Leucine aminopeptidases(LAPs)and its role in the process of growth and development,the Pseudomonas Cr13 LAP gene was cloned and characterized by bioinformatics in this study.Firstly,the core fragment of LAP was cloned.Secondly,the full-length DNA of LAP gene was cloned by two Genome Walking.Lastly,LAP gene was analyzed by bioinformatics software.The result showed that the full-length DNA of LAP gene was 1491 bp in Pseudomonas Cr13.Bioinformatics analysis indicated that the LAP gene was located in the mitochondrion.The LAP gene had a conserved cytoplasmic aminopeptidase motif NTDAEGRL.The Pseudomonas Cr13 LAP promoter(426 bp)contained the basic elements such as TATA-Box and CAAT-box and some potential cis-acting element.Cloning of full-length DNA laid a good foundation for further research on LAP gene function.Cloning and characterization of the promoter region of Pseudomonas sp.Cr13 LAP gene provided substantial basis for further research on the mechanism of regulation and expression of LAP gene.