title: Efficient expression of truncated recombinant cadmium-metallothionein gene of a ciliate, Tetrahymena tropicalis lahorensis in Escherichia coli.
abstract: Truncated recombinant metallothionein GST-fusion protein has been successfully expressed in Escherichia coli. The previously identified novel Cd-inducible metallothionein (TMCd1) gene from the locally isolated ciliate, Tetrahymena tropicalis lahorensis, was inserted into a pET-41a vector, in frame with a sequence encoding an N-terminal glutathione-S-transferase (GST) tail. Truncated recombinant GST fusion protein has been purified by affinity column chromatography using glutathione sepharose. After enzymatic cleavage of GST tail with enterokinase, the truncated TMCd1 MT shows molecular weight of 11.5 kDa, corresponding to the expected value. This is the first successful report of expression of cadmium metallothionein gene of a ciliate, T. t. lahorensis, reported from this part of the world, in E. coli. This study will further help in characterization of metallothionein protein of this ciliate.
authors.affiliation: School of Biological Sciences, University of the Punjab, Quaid-i-Azam Campus, Lahore, 54590, Pakistan.
authors.forename: Rukhsana Nighat
authors.initials: RN
authors.lastname: Shuja
authors.forename: Shuja Uddin Ahmad
authors.initials: SU
authors.lastname: Taimuri
authors.forename: Farah Rauf
authors.initials: FR
authors.lastname: Shakoori
authors.forename: Abdul Rauf
authors.initials: AR
authors.lastname: Shakoori
mesh_terms: D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000378:metabolism*; D015870:Gene Expression; D008668:Metallothionein / Q000737:chemistry / Q000235:genetics*; D008958:Models, Molecular; D016133:Polymerase Chain Reaction; D011993:Recombinant Fusion Proteins / Q000378:metabolism*; D013768:Tetrahymena / Q000235:genetics*
chemical_list: D011993:Recombinant Fusion Proteins; C004203:cadmium-binding protein; D008668:Metallothionein