Rathore, Dharmendar ; Nayak, Surendra K. ; Batra, Janendra K. (1997) Overproduction of fungal ribotoxin α -sarcin in Escherichia coli: generation of an active immunotoxin Gene, 190 (1). pp. 31-35. ISSN 0378-1119
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Official URL: http://linkinghub.elsevier.com/retrieve/pii/S03781...
Related URL: http://dx.doi.org/10.1016/S0378-1119(96)00696-8
Abstract
α-Sarcin is a ribonucleolytic protein secreted by the mold Aspergillus giganteus. DNA encoding a-sarcin was isolated from the host and cloned into T7 promoter based E. coli expression vectors. Using bacterial outer membrane protein A (OmpA) signal sequence, properly processed recombinant (re-) protein was secreted into the culture medium while in the absence of a signal sequence protein remained insoluble in the bacterial inclusion bodies. The re-α-sarcin was purified to homogeneity by simple chromatographic techniques both from the insoluble and soluble sources with respective yields of 40-50 μg/ml and 2-3 μg/ml. The re-ribotoxin was functionally as active as the native toxin and preserved its specificity. The re-α-sarcin was used in the construction of an active immunotoxin targeted at the human cancer cells overexpressing transferrin receptor (TFR).
Item Type: | Article |
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Source: | Copyright of this article belongs to Elsevier Science. |
Keywords: | Expression; Recombinant protein; Secretion; Immunoconjugate; Purification; Targeting; Aspergillus giganteus |
ID Code: | 13352 |
Deposited On: | 11 Nov 2010 08:23 |
Last Modified: | 17 May 2011 09:14 |
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