Molecular cloning of a phytase gene (phy M) from Pseudomonas syringae MOK1
J Cho, C Lee, S Kang, J Lee, H Lee, J Bok, J Woo… - Current …, 2005 - Springer
A phytase gene (phy M) was cloned from Pseudomonas syringae MOK1 by two steps of
degenerate PCR and inverse PCR. This gene consists of 1,287 nucleotides and encodes a
polypeptide of 428 amino acids with a deduced molecular mass of 46,652 kDa. Based on its
amino acid sequence, the Phy M shares the active site RHGXRXP and HD sequence motifs,
typically characterized by histidine acid phosphatases familly. Each phy M gene fragment
encoding mature Phy M with its own signal sequence (pEPSS) and without (pEPSM) was …
degenerate PCR and inverse PCR. This gene consists of 1,287 nucleotides and encodes a
polypeptide of 428 amino acids with a deduced molecular mass of 46,652 kDa. Based on its
amino acid sequence, the Phy M shares the active site RHGXRXP and HD sequence motifs,
typically characterized by histidine acid phosphatases familly. Each phy M gene fragment
encoding mature Phy M with its own signal sequence (pEPSS) and without (pEPSM) was …
Abstract
A phytase gene (phy M) was cloned from Pseudomonas syringae MOK1 by two steps of degenerate PCR and inverse PCR. This gene consists of 1,287 nucleotides and encodes a polypeptide of 428 amino acids with a deduced molecular mass of 46,652 kDa. Based on its amino acid sequence, the Phy M shares the active site RHGXRXP and HD sequence motifs, typically characterized by histidine acid phosphatases familly. Each phy M gene fragment encoding mature Phy M with its own signal sequence (pEPSS) and without (pEPSM) was subcloned into the E. coli BL21 (DE3) expression vector, pET22b (+). The enzyme activity in crude extracts of clone pEPSM was 2.514 Umg−1 of protein, and about 10-fold higher than that of clone pEPSS.
Springer
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