Cloning, Expression and Purification of Pseudomonas putida ATCC12633 Creatinase.

ABSTRACT: BACKGROUND:Pseudomonas putida (P. putida) ATCC12633 can produce creatinase. It is a microbial enzyme which degrades creatinine in bacteria and provides source of carbon and nitrogen. Also, this enzyme is used in the enzymatic measurement of creatinine concentration for diagnosis of renal and muscles functions and diseases. Our purpose was recombinant production of creatinase for using in clinical measurement of serum or urine creatinine. METHODS:A 1209bp of open reading frame of creatinase was amplified by PCR from P. putida ATCC12633 genome and cloned into pET28a expression vector which was digested using NheI and XhoI restriction enzymes. Cloning was confirmed by colony PCR, double digestion analysis and sequencing. Recombinant pET28a vector was transformed to Escherichia coli (E. coli)

SUBMITTER: Afshari E 

PROVIDER: S-EPMC5650733 | biostudies-literature | 2017 Oct-Dec

REPOSITORIES: biostudies-literature