Cloning of porcine neuron growth inhibitory factor (metallothionein III) cDNA and expression of the gene in Saccharomyces cerevisiae

Gene
S H WangL Y Lin

Abstract

Growth inhibitory factor (GIF), a member of the metallothionein (MT) family, is also known as MTIII. This protein distinguishes itself from other MT isoforms by exerting an inhibitory effect on cortical neuron growth instead of metal ion buffering. In this work, we cloned MTIII genes from a porcine brain cDNA library. Two species of clones were isolated that vary with respect to one nt in the coding sequence. This discrepancy results in the translation of two MTIII primary structures having a different amino acid at residue 46. Herein, both MTIII cDNAs were constructed into an expression vector and transformed into yeast cells, respectively. The yeast carrying either MTIII gene displayed a similar metal tolerance when cultured in a medium containing metal. The resistance to metal toxicity was attributed to the expression of MTIII gene which was confirmed by RNA and protein analyses. The characteristics of the protein stability, metal binding affinity and ultraviolet absorption spectrum of the yeast produced MTIII are also compared with those of MTII. The comparison reveals that both MTs have similar physical characteristics. Moreover, circular dichroism spectrum of Cd saturated MTIII was analyzed as well. Typical Cys-Cd bands f...Continue Reading

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Citations

Aug 26, 2006·Se pu = Chinese journal of chromatography·Weijuan ZhengZichun Hua
Apr 8, 2000·Critical Reviews in Biochemistry and Molecular Biology·A T MilesV Rodilla
Oct 6, 2004·Preparative Biochemistry & Biotechnology·Wei-Juan ZhengZi-Chun Hua
Jun 20, 2002·Protein Expression and Purification·Sue-Hong WangSimon J T Mao

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