Cloning and characterization of the Halobacillus trueperi betH gene, encoding the transport system for the compatible solute glycine betaine.
Halobacillus trueperi accumulates glycine betaine under condition of high osmolarity. A fragment of the glycine betaine transporter betH gene was obtained from the genome of H. trueperi with degenerate primers. Through Southern blot hybridization and inverse PCR, a 5.1 kb EcoRI fragment containing the complete betH gene was identified and subsequently sequenced. The betH gene was predicted to encode a 55.2 kDa protein (504 amino acid residues) with 12 transmembrane regions. BetH showed 56% identity to the OpuD of Bacillus subtilis which belongs to the betaine/carnitine/choline transporter (BCCT) family. Its putative promoter region was highly homologous to sigmaB-dependent promoter of B. subtilis. A 2.6 kb fragment containing the betH gene was cloned into pUC18 and transformed into the Escherichia coli MKH13. The accumulation of glycine betaine in transformed E. coli MKH13 bacteria was confirmed using 13C nuclear magnetic resonance spectroscopy.
Lu
Weidong
W
Department of Microbiology, College of Biological Sciences, China Agricultural University and Key Laboratory of Agro-Microbial Resources and Application, Ministry of Agriculture, Beijing 100094, PR China.
Zhao
Baisuo
B
Feng
Deqin
D
Yang
Susheng
S
D000595:Amino Acid Sequence; D001406:Bacillaceae / Q000235:genetics* / Q000378:metabolism; D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D001483:Base Sequence; D001622:Betaine / Q000378:metabolism*; D002247:Carbon Isotopes; D003001:Cloning, Molecular; D015246:Deoxyribonuclease EcoRI; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D009682:Magnetic Resonance Spectroscopy; D026901:Membrane Transport Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods; D017422:Sequence Analysis, DNA; D014882:Water-Electrolyte Balance
D001426:Bacterial Proteins; D002247:Carbon Isotopes; D026901:Membrane Transport Proteins; D001622:Betaine; D015246:Deoxyribonuclease EcoRI