Jung-Hwa Kang , Jong-Hee Lee , Je-Hyeon Park , Sung-Hoi Huh , In-Soo Kong
{"title":"拟态弧菌磷脂酶基因的克隆与鉴定","authors":"Jung-Hwa Kang , Jong-Hee Lee , Je-Hyeon Park , Sung-Hoi Huh , In-Soo Kong","doi":"10.1016/S0005-2760(98)00100-3","DOIUrl":null,"url":null,"abstract":"<div><p>The phospholipase gene <em>phl</em> was identified from <em>Vibrio mimicus</em> (ATCC33653) and sequenced. The entire open reading frame (ORF) was composed of 1410 nucleotides and encoding 470 amino acids. The <em>phl</em> was placed upstream of hemolysin gene (<em>vmh</em>A) with opposite direction of transcription. From the BLAST search program, the deduced amino acids sequence showed 74.4% identity with phospholipase gene (<em>lec</em>) from <em>V. cholerae</em> El Tor. The entire ORF of phospholipase gene was amplified by PCR and inserted into an <em>Escherichia coli</em> expression vector, pET22b(+) and introduced <em>E. coli</em> BL21(DE3). SDS–PAGE demonstrated that a protein corresponding to the phospholipase was overexpressed and migrated at a molecular mass of 53 kDa.</p></div>","PeriodicalId":100162,"journal":{"name":"Biochimica et Biophysica Acta (BBA) - Lipids and Lipid Metabolism","volume":null,"pages":null},"PeriodicalIF":0.0000,"publicationDate":"1998-10-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/S0005-2760(98)00100-3","citationCount":"26","resultStr":"{\"title\":\"Cloning and identification of a phospholipase gene from Vibrio mimicus\",\"authors\":\"Jung-Hwa Kang , Jong-Hee Lee , Je-Hyeon Park , Sung-Hoi Huh , In-Soo Kong\",\"doi\":\"10.1016/S0005-2760(98)00100-3\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<div><p>The phospholipase gene <em>phl</em> was identified from <em>Vibrio mimicus</em> (ATCC33653) and sequenced. The entire open reading frame (ORF) was composed of 1410 nucleotides and encoding 470 amino acids. The <em>phl</em> was placed upstream of hemolysin gene (<em>vmh</em>A) with opposite direction of transcription. From the BLAST search program, the deduced amino acids sequence showed 74.4% identity with phospholipase gene (<em>lec</em>) from <em>V. cholerae</em> El Tor. The entire ORF of phospholipase gene was amplified by PCR and inserted into an <em>Escherichia coli</em> expression vector, pET22b(+) and introduced <em>E. coli</em> BL21(DE3). SDS–PAGE demonstrated that a protein corresponding to the phospholipase was overexpressed and migrated at a molecular mass of 53 kDa.</p></div>\",\"PeriodicalId\":100162,\"journal\":{\"name\":\"Biochimica et Biophysica Acta (BBA) - Lipids and Lipid Metabolism\",\"volume\":null,\"pages\":null},\"PeriodicalIF\":0.0000,\"publicationDate\":\"1998-10-02\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"https://sci-hub-pdf.com/10.1016/S0005-2760(98)00100-3\",\"citationCount\":\"26\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Biochimica et Biophysica Acta (BBA) - Lipids and Lipid Metabolism\",\"FirstCategoryId\":\"1085\",\"ListUrlMain\":\"https://www.sciencedirect.com/science/article/pii/S0005276098001003\",\"RegionNum\":0,\"RegionCategory\":null,\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"\",\"JCRName\":\"\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Biochimica et Biophysica Acta (BBA) - Lipids and Lipid Metabolism","FirstCategoryId":"1085","ListUrlMain":"https://www.sciencedirect.com/science/article/pii/S0005276098001003","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"","JCRName":"","Score":null,"Total":0}
Cloning and identification of a phospholipase gene from Vibrio mimicus
The phospholipase gene phl was identified from Vibrio mimicus (ATCC33653) and sequenced. The entire open reading frame (ORF) was composed of 1410 nucleotides and encoding 470 amino acids. The phl was placed upstream of hemolysin gene (vmhA) with opposite direction of transcription. From the BLAST search program, the deduced amino acids sequence showed 74.4% identity with phospholipase gene (lec) from V. cholerae El Tor. The entire ORF of phospholipase gene was amplified by PCR and inserted into an Escherichia coli expression vector, pET22b(+) and introduced E. coli BL21(DE3). SDS–PAGE demonstrated that a protein corresponding to the phospholipase was overexpressed and migrated at a molecular mass of 53 kDa.