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Article

Expression, purification and characterization of a recombinant Tat47-57-Oct4 fusion protein in Pichia pastoris

  • Authors:
    • Haotian Wang
    • Xinmin Zhang
    • Ning Kong
    • Anhui Wei
    • Yanhong Zhang
    • Jie Ma
    • Yulai Zhou
    • Weiqun Yan
  • View Affiliations / Copyright

    Affiliations: School of Pharmaceutical Sciences of Jilin University, Changchun, Jilin 130021, P.R. China, College of Basic Medical Science of China Medical University, Shenyang, Liaoning 110001, P.R. China
  • Pages: 471-475
    |
    Published online on: December 11, 2013
       https://doi.org/10.3892/mmr.2013.1857
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Abstract

The transcription factor, Oct-4, is involved in the self-renewal of undifferentiated embryonic stem cells, and is also significant in the reprogramming process and in the development of tumors. In the present study, the fusion protein, Tat47‑57-Oct4, was secreted by the signal peptide of human serum albumin in Pichia pastoris under the control of alcohol oxidase promoter 1. The yield of recombinant Tat47‑57-Oct4 fusion protein was ~210 mg/l. Following pilot‑scale fermentation, Tat47‑57-Oct4 was purified by ammonium sulfate precipitation, Vivaflow 200 ultrafiltration and SP Sepharose fast flow chromatography in order to obtain 95.6% purity. Immunofluorescence analysis validated the ability of Tat47‑57-Oct4 to cross the cell membrane. The results demonstrated that the experimental procedure developed in the present study could produce large quantities of active Tat47‑57-Oct4 fusion protein from P. pastoris.
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1 

Takahashi K and Yamanaka S: Induction of pluripotent stem cells from mouse embryonic and adult fibroblast cultures by defined factors. Cell. 126:663–676. 2006. View Article : Google Scholar : PubMed/NCBI

2 

Vierbuchen T, Ostermeier A, Pang ZP, Kokubu Y, Südhof TC and Wernig M: Direct conversion of fibroblasts to functional neurons by defined factors. Nature. 463:1035–1041. 2010. View Article : Google Scholar : PubMed/NCBI

3 

Jiang B, Dong H, Li Q, Yu Y and Zhang Z, Zhang Y, Wang G and Zhang Z: Differentiation of reprogrammed mouse cardiac fibroblasts into functional cardiomyocytes. Cell Biochem Biophys. 66:309–318. 2013. View Article : Google Scholar : PubMed/NCBI

4 

Rosner MH, Vigano MA, Ozato K, Timmons PM, Poirier F, Rigby PW and Staudt LM: A POU-domain transcription factor in early stem cells and germ cells of the mammalian embryo. Nature. 345:686–692. 1990. View Article : Google Scholar : PubMed/NCBI

5 

Niwa H, Miyazaki J and Smith AG: Quantitative expression of Oct-3/4 defines differentiation, dedifferentiation or self-renewal of ES cells. Nat Genet. 24:372–376. 2000. View Article : Google Scholar : PubMed/NCBI

6 

Marión RM, Strati K, Li H, Murga M, Blanco R, Ortega S, Fernandez-Capetillo O, Serrano M and Blasco MA: A p53-mediated DNA damage response limits reprogramming to ensure iPS cell genomic integrity. Nature. 460:1149–1153. 2009.PubMed/NCBI

7 

Kim JB, Sebastiano V, Wu G, Araúzo-Bravo MJ, Sasse P, Gentile L, Ko K, Ruau D, Ehrich M, van den Boom D, et al: Oct4-induced pluripotency in adult neural stem cells. Cell. 136:411–419. 2009. View Article : Google Scholar : PubMed/NCBI

8 

Lam CS, Mistri TK, Foo YH, Sudhaharan T, Gan HT, Rodda D, Lim LH, Chou C, Robson P, Wohland T and Ahmed S: DNA-dependent Oct4-Sox2 interaction and diffusion properties characteristic of the pluripotent cell state revealed by fluorescence spectroscopy. Biochem J. 448:21–33. 2012. View Article : Google Scholar : PubMed/NCBI

9 

Moy P, Daikh Y, Pepinsky B, Thomas D, Fawell S and Barsoum J: Tat-mediated protein delivery can facilitate MHC class I presentation of antigens. Mol Biotechnol. 6:105–113. 1996. View Article : Google Scholar : PubMed/NCBI

10 

Nagahara H, Vocero-Akbani AM, Snyder EL, Ho A, Latham DG, Lissy NA, Becker-Hapak M, Ezhevsky SA and Dowdy SF: Transduction of full-length TAT fusion proteins into mammalian cells: TAT-p27Kip1 induces cell migration. Nat Med. 4:1449–1452. 1998. View Article : Google Scholar : PubMed/NCBI

11 

Schwarze SR, Ho A, Vocero-Akbani A and Dowdy SF: In vivo protein transduction: delivery of a biologically active protein into the mouse. Science. 285:1569–1572. 1999. View Article : Google Scholar : PubMed/NCBI

12 

Huang Z, Ji M, Peng Z, Huang S, Xiao Q, Li C, Zeng J, Gao M and Feng W: Purification of TAT-CC-HA protein under native condition, and its transduction analysis and biological effects on BCR-ABL positive cells. Biomed Pharmacother. 65:183–192. 2011. View Article : Google Scholar : PubMed/NCBI

13 

Park JS, Park SY, Cho HI, Sohn HJ and Kim TG: Enhanced induction of T cell immunity using dendritic cells pulsed with HIV Tat and HCMV-pp65 fusion protein in vitro. Immune Netw. 11:182–189. 2011. View Article : Google Scholar : PubMed/NCBI

14 

Zhao B, Wang Y, Zhang Y, Li Y, Zhang X, Xu Y, Chen L, Li C, Ju Y and Meng S: TAT-mediated gp96 transduction to APCs enhances gp96-induced antiviral and antitumor T cell responses. Vaccine. 31:545–552. 2013. View Article : Google Scholar : PubMed/NCBI

15 

Zhou H, Wu S, Joo JY, Zhu S, Han DW, Lin T, Trauger S, Bien G, Yao S, Zhu Y, et al: Generation of induced pluripotent stem cells using recombinant proteins. Cell Stem Cell. 4:381–384. 2009. View Article : Google Scholar : PubMed/NCBI

16 

Kim D, Kim CH, Moon JI, Chung YG, Chang MY, Han BS, Ko S, Yang E, Cha KY, Lanza R and Kim KS: Generation of human induced pluripotent stem cells by direct delivery of reprogramming proteins. Cell Stem Cell. 4:472–476. 2009. View Article : Google Scholar : PubMed/NCBI

17 

Sen Gupta C and Dighe RR: Hyperexpression of biologically active human chorionic gonadotropin using the methylotropic yeast, Pichia pastoris. J Mol Endocrinol. 22:273–283. 1999.

18 

Damaso MC, Almeida MS, Kurtenbach E, Martins OB, Pereira N Jr, Andrade CM and Albano RM: Optimized expression of a thermostable xylanase from Thermomyces lanuginosus in Pichia pastoris. Appl Environ Microbiol. 69:6064–6072. 2003. View Article : Google Scholar : PubMed/NCBI

19 

Murasugi A, Kido I, Kumai H and Asami Y: Efficient production of recombinant human pleiotrophin in yeast, Pichia pastoris. Biosci Biotechnol Biochem. 67:2288–2290. 2003. View Article : Google Scholar : PubMed/NCBI

20 

Laborde C, Chemardin P, Bigey F, Combarnous Y, Moulin G and Boze H: Overexpression of ovine leptin in Pichia pastoris: physiological yeast response to leptin production and characterization of the recombinant hormone. Yeast. 21:249–263. 2004.PubMed/NCBI

21 

Sreekrishna K, Brankamp RG, Kropp KE, Blankenship DT, Tsay JT, Smith PL, Wierschke JD, Subramaniam A and Birkenberger LA: Strategies for optimal synthesis and secretion of heterologous proteins in the methylotrophic yeast Pichia pastoris. Gene. 190:55–62. 1997. View Article : Google Scholar : PubMed/NCBI

22 

Bradford MM: A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 72:248–254. 1976. View Article : Google Scholar : PubMed/NCBI

23 

Sambrook J and Russell DW: SDS-polyacrylamide gel electrophoresis of proteins. CSH Protoc. 2006. View Article : Google Scholar

24 

Kim DW, Eum WS, Jang SH, Kim SY, Choi HS, Choi SH, An JJ, Lee SH, Lee KS, Han K, Kang TC, Won MH, Kang JH, Kwon OS, Cho SW, Kim TY, Park J and Choi SY: Transduced Tat-SOD fusion protein protects against ischemic brain injury. Mol Cells. 19:88–96. 2005.PubMed/NCBI

25 

Liu P, Liu X, Akf Liou EY, Xing J, Jing Z, Ji X, Liu X, Zhao H, Yan F, Chen J, Cao G and Luo Y: The Neuroprotective Mechanism of Erythropoietin-TAT Fusion Protein Against Neurodegeneration from Ischemic Brain Injury. CNS Neurol Disord Drug Targets. Aug 27–2013.(Epub ahead of print).

26 

Chengliang Zhou, Fengqing Xu, Chunhong Wang, Tao Liu, Xinrong Peng and Qijun Qian: Expression of human Oct4 and cell penetrating peptide fusion protein. Academic Journal of Second Military Medical University. 31:489–493. 2010.(In Chinese).

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Copy and paste a formatted citation
Spandidos Publications style
Wang H, Zhang X, Kong N, Wei A, Zhang Y, Ma J, Zhou Y and Yan W: Expression, purification and characterization of a recombinant Tat47-57-Oct4 fusion protein in Pichia pastoris. Mol Med Rep 9: 471-475, 2014.
APA
Wang, H., Zhang, X., Kong, N., Wei, A., Zhang, Y., Ma, J. ... Yan, W. (2014). Expression, purification and characterization of a recombinant Tat47-57-Oct4 fusion protein in Pichia pastoris. Molecular Medicine Reports, 9, 471-475. https://doi.org/10.3892/mmr.2013.1857
MLA
Wang, H., Zhang, X., Kong, N., Wei, A., Zhang, Y., Ma, J., Zhou, Y., Yan, W."Expression, purification and characterization of a recombinant Tat47-57-Oct4 fusion protein in Pichia pastoris". Molecular Medicine Reports 9.2 (2014): 471-475.
Chicago
Wang, H., Zhang, X., Kong, N., Wei, A., Zhang, Y., Ma, J., Zhou, Y., Yan, W."Expression, purification and characterization of a recombinant Tat47-57-Oct4 fusion protein in Pichia pastoris". Molecular Medicine Reports 9, no. 2 (2014): 471-475. https://doi.org/10.3892/mmr.2013.1857
Copy and paste a formatted citation
x
Spandidos Publications style
Wang H, Zhang X, Kong N, Wei A, Zhang Y, Ma J, Zhou Y and Yan W: Expression, purification and characterization of a recombinant Tat47-57-Oct4 fusion protein in Pichia pastoris. Mol Med Rep 9: 471-475, 2014.
APA
Wang, H., Zhang, X., Kong, N., Wei, A., Zhang, Y., Ma, J. ... Yan, W. (2014). Expression, purification and characterization of a recombinant Tat47-57-Oct4 fusion protein in Pichia pastoris. Molecular Medicine Reports, 9, 471-475. https://doi.org/10.3892/mmr.2013.1857
MLA
Wang, H., Zhang, X., Kong, N., Wei, A., Zhang, Y., Ma, J., Zhou, Y., Yan, W."Expression, purification and characterization of a recombinant Tat47-57-Oct4 fusion protein in Pichia pastoris". Molecular Medicine Reports 9.2 (2014): 471-475.
Chicago
Wang, H., Zhang, X., Kong, N., Wei, A., Zhang, Y., Ma, J., Zhou, Y., Yan, W."Expression, purification and characterization of a recombinant Tat47-57-Oct4 fusion protein in Pichia pastoris". Molecular Medicine Reports 9, no. 2 (2014): 471-475. https://doi.org/10.3892/mmr.2013.1857
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