| Title: | Codon optimization and expression of irisin in Pichia pastoris GS115 |
| Author(s): | Duan H; Wang H; Ma B; Jiang P; Tu P; Ni Z; Li X; Li M; Ma X; Wang B; Wu R; Li M; |
| Address: | "State Key Laboratory of Medicinal Chemical Biology, Nankai University, 300071 Tianjin, China; Key Laboratory for Bioactive Materials of the Ministry of Education, Institute of Molecular Biology, College of Life Science, Nankai University, 300071 Tianjin, China. Tianjin Children's Hospital, 300074 Tianjin, China. State Key Laboratory of Medicinal Chemical Biology, Nankai University, 300071 Tianjin, China. State Key Laboratory of Medicinal Chemical Biology, Nankai University, 300071 Tianjin, China; Key Laboratory for Bioactive Materials of the Ministry of Education, Institute of Molecular Biology, College of Life Science, Nankai University, 300071 Tianjin, China. Electronic address: mgl@nankai.edu.cn" |
| DOI: | 10.1016/j.ijbiomac.2015.04.030 |
| ISSN/ISBN: | 1879-0003 (Electronic) 0141-8130 (Linking) |
| Abstract: | "Irisin is a novel hormone which is related to many metabolic diseases. In order to illuminate the function and therapeutic effect of irisin, gaining active irisin is necessary. In this work, a codon-optimized irisin gene was designed according to Pichia pastoris synonymous codon usage bias and cloned into the pPIC9K expression vector. Sequencing result indicating that the sequence of irisin was consistent with the modified irisin and the irisin was in frame with alpha-factor secretion signal ATG. The plasmid pPIC9K-irisin was transformed into GS115 P. pastoris cells through electroporation. The positive transformants were screened on MD medium and analyzed by PCR. Five recombinant GS115/pPIC9K-irisin strains were obtained, but only one strain expressed irisin successfully. SDS-PAGE and Western blot were used to assess the expression level and purity of irisin. The irisin was also simply purified and the effect of pH value, methanol concentration and induction time on the production of irisin was investigated. The results showed that the best conditions of irisin expression were as follows: pH 6.0, 2.0% methanol and induction for 96 h. This work laid the basis for further investigation into the therapeutic and pharmacological effects of irisin, as well as development of irisin-based therapy" |
| Keywords: | "Amino Acid Sequence Base Sequence Cloning, Molecular *Codon Electroporation Fibronectins/biosynthesis/*genetics/isolation & purification Gene Expression Humans Hydrogen-Ion Concentration Mating Factor Methanol/metabolism/pharmacology Molecular Sequence Da;" |
| Notes: | "MedlineDuan, Huikun Wang, Haisong Ma, Baicheng Jiang, Pingzhe Tu, Peipei Ni, Zaizhong Li, Xiaodan Li, Miao Ma, Xiaofeng Wang, Bin Wu, Ri Li, Minggang eng Research Support, Non-U.S. Gov't Netherlands 2015/05/02 Int J Biol Macromol. 2015 Aug; 79:21-6. doi: 10.1016/j.ijbiomac.2015.04.030. Epub 2015 Apr 27" |
