Cloning, nucleotide sequence and expression of the gene encoding the cellulose-binding protein A (CBPA) of Eubacterium cellulosolvens 5.
The nucleotide sequence of the gene encoding the cellulose-binding protein A (CBPA) of Eubacterium cellulosolvens 5 was determined. The gene consists of an open reading frame of 3453 nucleotides and encodes a protein of 1151 amino acids with a molecular mass of 126408 Da. The deduced amino acid sequence of CBPA contained one domain highly similar to a catalytic domain of glycosyl hydrolases belonging to family 9, two linker-like domains and four domains of unknown function. Among the four domains of unknown function, the domains 1 and 2 region had significant homology in amino acid sequence with the cellulose-binding domains in the family 9 glycosyl hydrolases. The cloned gene was inserted into an expression vector, pBAD-TOPO, and expressed in Escherichia coli as a fused protein. The fused protein was detected by immunoblotting using antiserum against CBPA.
Toyoda
Atsushi
A
School of Agriculture, Ibaraki University, Ami, 300-0393, Ibaraki, Japan. atoyoda@acs.ipc.ibaraki.ac.jp
Minato
Hajime
H
D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics*; D001483:Base Sequence; D002352:Carrier Proteins / Q000737:chemistry / Q000235:genetics*; D002482:Cellulose / Q000378:metabolism; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics; D005051:Eubacterium / Q000737:chemistry / Q000235:genetics*; D005798:Genes, Bacterial; D005822:Genetic Vectors; D008969:Molecular Sequence Data; D008970:Molecular Weight; D016415:Sequence Alignment
D001426:Bacterial Proteins; D002352:Carrier Proteins; D002482:Cellulose