Sekizawa Y, Kubo T, Kobayashi H, Nakajima T, Natori S
Research Laboratory, Zenyaku Kogyo Co., Ltd., Nerima-ku, Tokyo, Japan.
Gene. 1997 May 20;191(1):97-102. doi: 10.1016/s0378-1119(97)00047-4.
Lysenin, which causes contraction of rat vascular smooth muscle, is a protein that was isolated from the earthworm Eisenia foetida. A cDNA encoding lysenin was isolated by use of a partial cDNA probe that had been generated by the PCR with a primer designed by reference to an internal peptide sequence of lysenin. This clone had an ORF encoding 297 amino acid residues. The amino acid sequence deduced from the cDNA revealed the absence of any significant homology to those of previously characterized vasoactive substances. The recombinant lysenin was produced in Escherichia coli. This protein and native lysenin isolated from the earthworm had similar contractive activities when tested on rat aorta. Northern blot analysis of the RNA from various tissues of the earthworm indicated that lysenin is produced by the coelomocytes.
引起大鼠血管平滑肌收缩的蚯蚓溶细胞素是一种从赤子爱胜蚓中分离出的蛋白质。利用通过PCR产生的部分cDNA探针分离出了编码蚯蚓溶细胞素的cDNA,该探针是使用参照蚯蚓溶细胞素内部肽序列设计的引物制备的。该克隆有一个编码297个氨基酸残基的开放阅读框。从cDNA推导的氨基酸序列显示与先前表征的血管活性物质的氨基酸序列没有任何显著同源性。重组蚯蚓溶细胞素在大肠杆菌中产生。当在大鼠主动脉上进行测试时,这种蛋白质和从蚯蚓中分离出的天然蚯蚓溶细胞素具有相似的收缩活性。对蚯蚓各种组织的RNA进行的Northern印迹分析表明,蚯蚓溶细胞素是由体腔细胞产生的。