TY - JOUR
T1 - Production of human serum transferrin in Escherichia coli
AU - Ikeda, Richard A.
AU - Bowman, Barbara H.
AU - Yang, Funmei
AU - Lokey, Laurie K.
PY - 1992/8/15
Y1 - 1992/8/15
N2 - Transferrin (Tf) crystals diffract to only medium resolution. The mediocre quality of the crystals may be due to two factors: (1) the genetic variations naturally present in the primary sequence ofTf, and (2) the glycosylation of the protein. To control genetic variations and glycosylation of samples of Tf, it would be desirable to express the Tf gene from a recombinant clone. Additionally, expression of Tf from a clone would allow for manipulation of the structure of Tf. The cDNA encoding Tf has been cloned into the pL-based expression vector, pRE1, and the T7-based expression vectors, pRSETA and pET11A. The tf expression plasmids, pTF-SSn and pTF-ESn, based on the T7 expression vectors, efficiently produce a 76-kDa protein that is approximately the same size as deglycosylated Tf, cross reacts with anti-Tf antibodies, and matches the deduced N-terminal amino acid sequence. Expression of Tf in Escherichia coli will allow the production of genetically pure, unglycosylated protein.
AB - Transferrin (Tf) crystals diffract to only medium resolution. The mediocre quality of the crystals may be due to two factors: (1) the genetic variations naturally present in the primary sequence ofTf, and (2) the glycosylation of the protein. To control genetic variations and glycosylation of samples of Tf, it would be desirable to express the Tf gene from a recombinant clone. Additionally, expression of Tf from a clone would allow for manipulation of the structure of Tf. The cDNA encoding Tf has been cloned into the pL-based expression vector, pRE1, and the T7-based expression vectors, pRSETA and pET11A. The tf expression plasmids, pTF-SSn and pTF-ESn, based on the T7 expression vectors, efficiently produce a 76-kDa protein that is approximately the same size as deglycosylated Tf, cross reacts with anti-Tf antibodies, and matches the deduced N-terminal amino acid sequence. Expression of Tf in Escherichia coli will allow the production of genetically pure, unglycosylated protein.
KW - Crystallization
KW - iron-binding protein
KW - phage T7 vectors
KW - protein sequencing
KW - recombinant DNA
KW - site-directed mutagenesis
KW - Western detection
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U2 - 10.1016/0378-1119(92)90737-A
DO - 10.1016/0378-1119(92)90737-A
M3 - Article
C2 - 1639274
AN - SCOPUS:0026651206
VL - 117
SP - 265
EP - 269
JO - Gene
JF - Gene
SN - 0378-1119
IS - 2
ER -