CLONING AND EXPRESSION IN ESCHERICHIA-COLI OF THE GENE FOR EXTRACELLULAR PHOSPHOLIPASE-A1 FROM SERRATIA-LIQUEFACIENS
CLONING AND EXPRESSION IN ESCHERICHIA-COLI OF THE GENE FOR EXTRACELLULAR PHOSPHOLIPASE-A1 FROM SERRATIA-LIQUEFACIENS
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DOI：
10.1128/jb.170.12.5855-5862.1988
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发表时间：
1988-12-01
影响因子：
3.2
通讯作者：
MOLIN, S
中科院分区：
文献类型：
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作者：
GIVSKOV, M;OLSEN, L;MOLIN, S
从液化沙雷氏菌基因组文库中分离到一个含有双基因操纵子的克隆DNA片段，并在大肠杆菌中表达。其中一个基因产物被鉴定为磷脂酶A1，该酶被发现从液化葡萄球菌和大肠杆菌分泌到外部环境。对这两个基因进行了测序，并建立了DNA序列中的开放阅读框与体外表达的多肽之间的关系。该磷脂酶多肽的长度为319个氨基酸。在编码序列的氨基末端有大约20个疏水氨基酸，但与一致的信号肽不同的是，没有碱性残基。第二个多肽的长度为227个氨基酸。研究发现，磷脂酶基因在大肠杆菌和液化葡萄球菌中的表达均受生长阶段调控(晚期表达)。
From a genomic library of Serratia liquefaciens, a cloned DNA fragment comprising a two-gene operon was isolated and expressed in Escherichia coli. One of the gene products was identified as a phospholipase A1, and the enzyme was found to be excreted to the outer environment from S. liquefaciens as well as from E. coli. Both genes were sequenced, and the relationship between open reading frames in the DNA sequence and in vitro-expressed polypeptides was established. The length of the phospholipase polypeptide was found to be 319 amino acids. In the amino-terminal end of the coding sequence was a stretch of about 20 hydrophobic amino acids, but, in contrast to consensus signal peptides, no basic residues were present. The length of the second polypeptide was 227 amino acids. It was found that expression of the phsopholipase gene in both E. coli and S. liquefaciens was growth phase regulated (late expression).