Kempf B, Bremer E
Max-Planck-Institut für Terrestrische Mikrobiologie, Karl-von-Frisch Strasse, Marburg, Germany.
FEMS Microbiol Lett. 1996 Aug 1;141(2-3):129-37. doi: 10.1111/j.1574-6968.1996.tb08374.x.
The nucleotide sequence of a new Bacillus subtilis gene (amhX) was determined that encodes a protein (AmhX) with strong sequence identity to amidohydrolases from both plant and bacterial species and a carboxypeptidase from the archaeon Sulfolobus sulfataricus. The amhX gene encodes a hydrophilic polypeptide of 383 amino acids with a molecular mass of 41.5 kDa. The amhX gene was overexpressed in E. coli by using the T7 RNA polymerase/promoter system and the transcription initiation sites for the amhX mRNAs in B. subtilis were determined by primer extension analysis. Chromosomal amhX mutations were constructed by marker replacement and the amhX gene was positioned at 25 degrees on the genetic and physical map of the B. subtilis chromosome.
测定了一种新的枯草芽孢杆菌基因（amhX）的核苷酸序列，该基因编码一种蛋白质（AmhX），它与来自植物和细菌物种的酰胺水解酶以及古菌嗜热栖热菌的一种羧肽酶具有很强的序列同一性。amhX基因编码一个由383个氨基酸组成的亲水性多肽，分子量为41.5 kDa。通过使用T7 RNA聚合酶/启动子系统在大肠杆菌中对amhX基因进行了过表达，并通过引物延伸分析确定了枯草芽孢杆菌中amhX mRNA的转录起始位点。通过标记替换构建了染色体amhX突变体，并将amhX基因定位在枯草芽孢杆菌染色体的遗传和物理图谱上25度处。
