[The cloning and expression of salmon calcitonin gene in Escherichia coli].
The DNA fragment encoding salmon calcitonin was separated from the salmon DNA with PCR method. It was recombined into the fusion expression vector (pGEX-2T) which included a Glutathion-S-Transferase (GST) gene and was transformed into E. coli JM109 and then expression was induced with IPTG. The fusion protein (GST-sCT) accounted for 38%-40% of the total cellular protein and was purified from the soluble expression products by glutathion sepharose 4B affinity chromatography. The purity of GST-sCT is about 80% and it showed a positive immunological reaction in calcitonin enzyme immunoassay.
Di
X
X
Institute of Basic Medical Sciences, CAMS, Beijing.
Chen
S
S
Song
F
F
Zhang
J
J
Chen
W
W
D000595:Amino Acid Sequence; D001483:Base Sequence; D002116:Calcitonin / Q000096:biosynthesis* / Q000235:genetics; D003001:Cloning, Molecular; D004247:DNA / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics*; D015870:Gene Expression; D005982:Glutathione Transferase / Q000096:biosynthesis* / Q000235:genetics; D008969:Molecular Sequence Data; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis* / Q000235:genetics; D014170:Transformation, Genetic
D011993:Recombinant Fusion Proteins; C028815:salmon calcitonin; D002116:Calcitonin; D004247:DNA; D005982:Glutathione Transferase