Cloning and sequencing of a new holin-encoding gene of Bacillus licheniformis.
A Bacillus licheniformis DNA fragment which exhibits homology with the upstream region of the cell-wall hydrolase-encoding gene, cwlL, was cloned into Escherichia coli (Ec). Nucleotide sequencing indicated that there are two open reading frames (tentatively designated as xpaG1 and xpaG2) which encode polypeptides of 89 and 88 amino acids (aa) (10044 and 9764 Da, respectively). Ec cells harboring two compatible plasmids (pMWB1 and pHSGKH) containing the Bacillus subtilis cell-wall hydrolase-encoding gene, cwlA, and xpaG1-G2, respectively, exhibited higher extra-cellular cell-wall hydrolase activity than did cells harboring pMWB1 and a control plasmid, pHSG398. The aa sequence homology of XpaG2 with other polypeptides indicated that xpaG2 is a holin-encoding gene. Moreover, Ec C600 harboring a plasmid containing xpaG1-xpaG2 led to leakage of beta-galactosidase into the extracellular fraction.
Kyogoku
K
K
Department of Applied Biology, Shinshu University, Nagano, Japan.
Sekiguchi
J
J
D000581:Amidohydrolases / Q000235:genetics / Q000378:metabolism; D000595:Amino Acid Sequence; D001407:Bacillus / Q000201:enzymology / Q000235:genetics*; D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D001435:Bacteriophages / Q000235:genetics / Q000378:metabolism; D001483:Base Sequence; D002455:Cell Division; D002473:Cell Wall / Q000378:metabolism; D003001:Cloning, Molecular; D005798:Genes, Bacterial; D008969:Molecular Sequence Data; D009238:N-Acetylmuramoyl-L-alanine Amidase / Q000737:chemistry / Q000235:genetics; D010957:Plasmids; D017433:Protein Structure, Secondary; D017421:Sequence Analysis; D017385:Sequence Homology; D001616:beta-Galactosidase / Q000378:metabolism
D001426:Bacterial Proteins; D001616:beta-Galactosidase; D000581:Amidohydrolases; C098606:XpaG1 protein, Bacillus licheniformis; C098608:XpaG2 protein, Bacillus licheniformis; C084313:CwlL protein, Bacillus licheniformis; D009238:N-Acetylmuramoyl-L-alanine Amidase