Frontiers in Biology >

2007 , Vol. 2 >Issue 2: 170 - 175

DOI: https://doi.org/10.1007/s11515-007-0023-5

Cloning and expressing a recombinant human tissue inhibitor of metalloproteinase-2 (TIMP-2) in methylotrophic yeast Pichia pastoris and its characterizations

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  • 1.Insitute of Biotechnology, Academy of Military Medical Sciences, Beijing 100071, China; Biology Department, Langfang Normal College, Langfang 065000, China; College of Food Science, Southwest Agricultural University, Chongqing 400716, China; 2.Insitute of Biotechnology, Academy of Military Medical Sciences, Beijing 100071, China; 3.College of Food Science, Southwest Agricultural University, Chongqing 400716, China

Published date: 05 Jun 2007

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Abstract

To obtain human tissue inhibitor of metalloproteinase-2 (TIMP-2) cDNA and the secretory expression of TIMP-2 gene in Pichia pastoris, we designed and synthesized a 618 base pairs artificial gene coding for the TIMP-2 with a computer-aided design method using a standard chemical synthesis technique, which was composed of frequently used codons in the highly expressed Pichia pastoris genes. Then the synthetic gene encoding TIMP-2 was checked by means of dideoxynucleotide sequencing. The verified gene of TIMP-2 was cloned to the Escherichia coli-yeast shuttle vector of pPIC9 to construct a recombinant plasmid pPIC9-T2. The plasmid was transformed into GS115 cells of the methylotrophic yeast, Pichia pastoris by electroporation, and we got the expression cell through phenotype selection and induction with methanol. Separation, purification, and bioactivity analysis of the expressed products were performed.

Cite this article

YAN Xunyou, ZHAO Hongliang, XUE Chong, LIU Zhimin, ZHANG Weiguang . Cloning and expressing a recombinant human tissue inhibitor of metalloproteinase-2 (TIMP-2) in methylotrophic yeast Pichia pastoris and its characterizations[J]. Frontiers in Biology, 2007 , 2(2) : 170 -175 . DOI: 10.1007/s11515-007-0023-5

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