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Protein Expression & Purification Technologies

Protein expression and purification can be very challenging because there is no single approach that is appropriate for every protein or downstream application.

Successful expression can be affected by each individual protein’s solubility, toxicity, need for post-translational modifications or its intrinsic ability to fold. Additionally, the application for which the protein is ultimately to be used may dictate how the protein needs to be expressed or purified. It is therefore important to have a flexible set of tools and methods that can be applied to each unique project.

Each NEB expression technology offers different advantages enabling you to choose the strategy that best suits your protein expression and purification needs. Many of the expression vectors share a compatible polylinker, enabling the gene of interest to be easily shuffled between systems. Various applications along with the recommended NEB kit can be found by clicking the sidebar tab.

Protein Expression & Purification Technologies includes these subcategories:

Cell-Free Expression
E. coli
Magnetic Matrices & Racks
Yeast

FAQs for Protein Expression & Purification Technologies

Protocols for Protein Expression & Purification Technologies

    Publications related to Protein Expression & Purification Technologies

  1. Tait, A.R. and Straus, S.K. 2011. Overexpression and purification of U24 from human herpesvirus type-6 in E. coli: unconvential use of oxidizing environments with a maltose binding protein-hexahistidine dual tag to enhance membrane protein yield Microbial Cell Factories. 10, PubMedID: 21130169, DOI:
  2. Ritter, A. 2011. Next-gen expression systems . 35, PubMedID: , DOI:
  3. Whitney, M.A. et al. 2011. Fluorescent peptides highlight peripheral nerves during surgery in mice Nat. Biotechnol. . 6, PubMedID: 21297616, DOI:
  4. Salinas, G., Pellizza, L., Margenat, M., et al. 2011. Tuned Escherichia coli as a host for the expression of disulfide-rich proteins Biotechnology Journal. 6, PubMedID: 21567960, DOI:
  5. Correa, A. and Oppezzo, P. 2011. Tuning different expression parameters to achieve soluble recombinant proteins in E. coli: Advantages of high-throughput screening Biotechnology Journal. 6, PubMedID: 21567962, DOI:
  6. Zhi, M. et al. 2011. Screening of specific binding peptide targeting blood vessel of human esophageal cancer in vivo in mice Chin. Med. J. . 124, PubMedID: 21362285, DOI:
  7. Ganatra, M.B., Rainauskas, S., Hong J.M., Taylor, T.E., Denson, J.P.M., Esposito, D., Read, J.D., Schmeisser, H., Zoon, K.C., Hartley, J.L. and Taron, C.H. 2011. A set of aspartyl protease-deficient strains for improved expression of heterologous proteins in Kluyveromyces lactis FEMS Yeast Res.. 11, PubMedID: 21166768, DOI:
  8. Sawada, R. et al. 2011. A phage-targeting startegy for the design of spatiotemporal drug delivery from grafted matrices Fibrogenesis Tissue Repair. 4, PubMedID: 17641242, DOI:
  9. Sun, Z. et al. 2013. High-resolution enzymatic mapping of 5-hydroxymethylcytosine in mouse embryonic stem cells Cell Rep.. , PubMedID: 23352666, DOI:
  10. Zahid, M. et al. . Identification of a cardiac specific protein transduction domain by in vivo biopanning using a M13 phage peptide display library in mice PLoS One. 5, PubMedID: 20808875, DOI:
  11. Vernet, E., Kotzsch, A., Voldborg, B., et al. . Screening of genetic parameters for soluble protein expression in Escherichia coli Protein Expression and Purification. 77, PubMedID: 21130169, DOI:
  12. Ohashi, H., Kanamori, T., Shimizu, Y. and Ueda, T. 2010. A highly controllable reconstituted cell-free system--a breakthrough in protein synthesis research Curr. Pharm. Biotechnol.. 11, PubMedID: 20210745, DOI:
  13. Zhao, S. et al. 2010. Novel peptide ligands that bind specifically to mouse embryonic stem cells Peptides. 31, PubMedID: 20713104, DOI:
  14. Sun, X. et al. 2010. Phage display-derived peptides for osteosarcoma imaging Clin. Cancer Res.. 16, PubMedID: 20570932, DOI:
  15. Zhao, W. et al. 2010. Isolation and initial application of a novel peptide that specifically recognizes the neural stem cells derived from rhesus monkey embryonic stem cells J. Biomol. Screen.. 15, PubMedID: 20508256, DOI:
  16. Yoon, S.H., Kim, S.K., and Kim J.F. 2010. Secretory production of recombinant proteins in Escherichia coli Recent Patents on Biotechnology. 4, PubMedID: 20201800, DOI:
  17. Wu, D. et al. 2010. Anti-tumor effects of a novel chimeric peptide on S180 and H22 xenografts bearing nude mice Peptides . 31, PubMedID: 20132854, DOI:
  18. Wu, C.C. et al. 2010. Identification of a new peptide for fibrosarcoma tumor targeting and imaging in vivo J. Biomed. Biotechnol. . , PubMedID: 21151669, DOI:
  19. Seow, Y. et al. 2010. Identification of a novel muscle targeting peptide in mdx mice Peptides. 31, PubMedID: 20621144, DOI:
  20. Welch, B.D. et al. 2010. Design of a potent D-peptide HIV-1 entry inhibitor with a strong barrier to resistance J. Virol.. 84, PubMedID: 20719956, DOI:
  21. Berkmen, M. 2012. Production of disulfide-bonded proteins in Escherichia coli Protein Expression and Purification. , PubMedID: 22085722, DOI:
  22. Berkmen, M. 2012. Oxidative folding of proteins in Escherichia coli BioPharm International. , PubMedID: , DOI:
  23. Tzertzinis, G., Schildkraut, E., and Schildkraut, I. 2012. Substrate cooperativity in marine luciferases PLoS One. 7, PubMedID: 22768230, DOI:
  24. Lobstein, J., Emrich, C.A., Jeans, C., et al. 2012. SHuffle, a novel Escherichia coli protein expression strain capable of correctly folding disulfide bonded proteins in its cytoplasm Microbial Cell Factories. 11, PubMedID: 22569138, DOI:
  25. Samuelson, J.C., Causey, T.B., and Berkmen, M. 2012. Disulfide-bonded protein expression in E. coli Genetic Engineering & Biotechnology News. 32(3), PubMedID: , DOI:
  26. Lamppa, J.W., Tanyos, S.A., and Griswold, K.E. 2012. Engineering Escherichia coli for soluble expression and single step purification of active human lysozyme Journal of Biotechnology. , PubMedID: 23220215, DOI: S0168-S01656
  27. Zitzman, S. et al. 2007. Identification and evaluation of a new tumor cell-binding peptide J. Nucl. Med.. 48, PubMedID: 17504878, DOI:
  28. Schmidt, A. et al. 2007. Selecting targeting of adenoviral vectors to neural precursor cells in the hippocampus of adult mice: new prospects for in situ gene therapy Stem Cells. 25, PubMedID: 17641242, DOI:
  29. Wan, X.M. et al. 2009. Identification of nose-to-brain homing peptide through phage display Peptides. 30, PubMedID: 343-350, DOI:
  30. de Marco, A. 2009. Strategies for successful recombinant expression of disulfide bond-dependent proteins in Escherichia coli Microbial Cell Factories. 8, PubMedID: 19442264, DOI:
  31. Mueller, J. et al. 2009. Targeting of tumor blood vessels: a phage-displayed tumor-homing peptide specifically binds to matrix metalloproteinase-2-processed collagen IV and blocks angiogenesis in vivo Mol. Cancer Res.. 7, PubMedID: 19584266, DOI:
  32. Nothelfer, E.M. et al. 2009. Identification and characterization of a peptide with affinity to head and neck cancer J. Nucl. Med.. 50, PubMedID: 19223422, DOI:
  33. Shimizu, Y., Inoue, A., Tomari, Y., Suzuki, T., Yokogawa, T., Nishikawa, K. and Ueda, T. 2001. Cell-free translation reconstituted with purified components Nat. Biotech.. 19, PubMedID: 11479568, DOI:
  34. Dorsch, S., et al. 2001. The VP1-unique region of parvovirus B19: amino acid variability and antigenic stability J. Gen. Virol. . 82 (pt 1) , PubMedID: 11125172, DOI:
  35. Wu, J.W., et al. 2001. Crystal structure of a phosphorylated Smad2. Recognition of phosphoserine by the MH2 domain and insights on Smad function in TGF-beta signaling Mol Cell.. 8, PubMedID: 11779503, DOI:
  36. Hanzawa, H., et al. 2001. In vitro assembly of phytochrome B apoprotein with synthetic analogs of the phytochrome chromophore Proc. Natl. Acad. Sci. USA . 98 , PubMedID: 11248126, DOI:
  37. Midon, M., Schäfer, P., Pingoud, A., Ghosh, M., Moon, A.F., Cuneo, M.J., London, R.E. and Meiss, G. 2011. Mutational and biochemical analysis of the DNA-entry nuclease EndA from Streptococcus pneumoniae Nucleic Acids Res. . 39, PubMedID: 20846957, DOI:
  38. Iizuka, R., Yamagishi-Shirasaki, M. and Funatsu, T. 2011. Kinetic study of de novo chromophore maturation of fluorescent proteins Anal. Biochem. .  Apr 1. , PubMedID: 21459075 , DOI:
  39. Feng, Z., et al. 2011. Optimization of Medium Composition for Production of Recombinant Calf Chymosin from Kluyveromyces lactis in Submerged Fermentation J. of Northeast Agricultural University. 18, PubMedID: , DOI:
  40. Rocha S.N., et al. 2011. Heterologous expression of a thermophilic esterase in Kluyveromyces yeasts Appl. Microbiol. Biotechnol.. 89, PubMedID: 20862582, DOI:
  41. Yamamoto, S., Izumiya, H., Mitobe, J., Morita, M., Arakawa, E., Ohnishi, M. and Watanabe, H. 2011. Identification of a chitin-induced small RNA that regulates translation of the tfoX gene, encoding a positive regulator of natural competence in Vibrio cholerae J Bacteriol. . 193, PubMedID: 21317321, DOI:
  42. De Masi, F., Grove, C.A., Vedenko, A., Alibés, A., Gisselbrecht, S.S., Serrano, L., Bulyk, M.L. and Walhout, A.J. 2011. Using a structural and logics systems approach to infer bHLH-DNA binding specificity determinants Nucleic Acids Res. . 18-Feb , PubMedID: 21335608 , DOI:
  43. Handa, Y., Inaho, N. and Nameki, N. 2011. YaeJ is a novel ribosome-associated protein inEscherichia coli that can hydrolyze peptidyl-tRNA on stalled ribosomes Nucleic Acids Res.  . 39, PubMedID: 21051357, DOI:
  44. Panayiotou, C., Solaroli, N., Xu, Y., Johansson, M. and Karlsson, A. 2011. The characterization of human adenylate kinases 7 and 8 demonstrates differences in kinetic parameters and structural organization among the family of adenylate kinase isoenzymes Biochem J.  . 2433 , PubMedID: 21080915, DOI:
  45. Lamichhane, T.N., Abeydeera, N.D., Duc, A.C., Cunningham, P.R. and Chow, C.S 2011. Selection of peptides targeting helix 31 of bacterial 16S ribosomal RNA by screening M13 phage-display libraries Molecules . 16, PubMedID: 21278676, DOI:
  46. Ganatra, M.B., Rainauskas, S., Hong, J.M., Taylor, T.E., Denson, J.P.M., Esposito, D., Read, J.D., Schmeisser, H., Zoon, K.C., Hartley, J.L. and Taron, C.H. 2011. A set of aspartyl protease-deficient strains for improved expression of heterologous proteins in Kluyveromyces lactis FEMS Yeast Res.. 11, PubMedID: 21166768, DOI:
  47. Rosner, K., Kasprzak, M.F., Horenstein, A.C., Thurston, H.L., Abrams, J., Kerwin, L.Y., Mehregan, D.A. and Mehregan, D.R. 2011. Engineering a waste management enzyme to overcome cancer resistance to apoptosis: adding DNase1 to the anti-cancer toolbox Cancer Gene Ther. . 18, PubMedID: 21233855, DOI:
  48. Feng, Z., et al. 2011. Disruption of PMR1 in Kluyveromyces lactis improves secretion of calf prochymosin J. Sci. Food Agric.. 15, PubMedID: , DOI:
  49. Narayan, V., Pion, E., Landré, V., Müller, P. and Ball, K.L. 2011. Docking-dependent ubiquitination of the interferon regulatory factor-1 tumor suppressor protein by the ubiquitin ligase CHIP J. Biol. Chem. . 286, PubMedID: 20947504, DOI:
  50. Zhou, X., et al. 2011. Expression of heparin sulfate sulfotransferases in Kluyveromyces lactisand preparation of PAPS Glycobiology Advance Access. , PubMedID: , DOI:
  51. Wojciechowski CL, Cardia JP, and Kantrowitz ER. 2002. Alkaline phosphatase from the hyperthermophilic bacterium T. maritima requires cobalt for activity Protein Sci. 4, PubMedID: 11910033, DOI:
  52. Nakano, S., et al. 2002. Multiple pathways of Spx (YjbD) proteolysis in Bacillus subtilis J. Bacteriol. 184, PubMedID: 12057962, DOI:
  53. Singleton, S. F., et al. 2002. Intein-mediated affinity-fusion purification of the Escherichia coliRecA protein Protein Expr. Purif. . 26, PubMedID: , DOI:
  54. Moolenaar GF, et al. 2002. Cho, a second endonuclease involved in Escherichia coli nucleotide excision repair Proc. Natl. Acad. Sci. US. 99, PubMedID: 11818552, DOI:
  55. Camarero, J.A., et al. 2002. Autoregulation of a bacterial sigma factor explored by using segmental isotopic labeling and NMR Proc. Natl. Acad. Sci. USA. 99, PubMedID: 12084914, DOI:
  56. Wojciak, J.M., et al. 2002. Arm-site binding by lambda -integrase: solution structure and functional characterization of its amino-terminal domain Proc. Natl. Acad. Sci. USA . 99, PubMedID: 11904406, DOI:
  57. Zhong, Q., et al. 2002. Endosomal localization and function of sorting Proc. Natl. Acad. Sci. USA. 99, PubMedID: , DOI:
  58. Humphries, H. E., Christodoulides, M. and Heckels, J. E. 2002. Expression of the class 1 outer-membrane protein of Neisseria meningitidis in Escherichia coli and purification using a self-cleavable affinity tag Protein Expr. Purif. . 26, PubMedID: , DOI:
  59. Zuberek, J., et al. 2003. Phosphorylation of eIF4E attenuates its interaction with mRNA 5' cap analogs by electrostatic repulsion: intein-mediated protein ligation strategy to obtain phosphorylated protein RNA. 9, PubMedID: 12554876, DOI:
  60. Hong, S.H., and Maret, W. 2003. A fluorescence resonance energy transfer sensor for the beta-domain of metallothionein Proc. Natl. Acad. Sci. USA. 100, PubMedID: 12618543, DOI:
  61. Scheibner, K.A., Zhang, Z. and Cole, P.A. 2003. Merging fluorescence resonance energy transfer and expressed protein ligation to analyze protein-protein interactions Anal. Biochem. . 317, PubMedID: 12758261, DOI:
  62. Wuebbens, M. M. and Rajagopalan, K. V. 2003. Mechanistic and mutational studies of Escherichia coli molybdopterin synthase clarify the final step of molybdopterin biosynthesis J. Biol. Chem. . 278, PubMedID: 12571226, DOI:
  63. Bonsager, B.C., et al. 2003. Purification and characterization of the beta-trefoil fold protein barley alpha-amylase/subtilisin inhibitor overexpressed in Escherichia coli Protein Expr. Purif. . 2, PubMedID: 12880767, DOI:
  64. Sinars, C.R., et al. 2003. Structure of the large FK506-binding protein FKBP51, an Hsp90-binding protein and a component of steroid receptor complexes Proc. Natl. Acad. Sci. USA. 100, PubMedID: 12538866, DOI:
  65. Zhang, Z., et al. 2003. The role of C-terminal tyrosine phosphorylation in the regulation of SHP-1 explored via expressed protein ligation J. Biol. Chem.. 278, PubMedID: 12468540, DOI:
  66. Rak, A., et al. 2003. Structure of Rab GDP-dissociation inhibitor in complex with prenylated YPT1 GTPase Science. 302, PubMedID: 14576435, DOI:
  67. David, R., et al. 2003. Semisynthesis and application of carboxyfluorescein-labelled biologically active human interleukin-8 Biol. Chem.. 384, PubMedID: 14719805, DOI:
  68. Schaffer, L., et al. 2003. Assembly of high-affinity insulin receptor agonists and antagonists from peptide building blocks Proc. Natl. Acad. Sci. USA. 100, PubMedID: 12684539, DOI:
  69. Salazar, J.C., et al. 2003. Coevolution of an aminoacyl-tRNA synthetase with its tRNA substrates Proc. Natl. Acad. Sci. USA. 100, PubMedID: 14615592, DOI:
  70. Walters, K.J., et al. 2003. DNA -repair protein hHR23a alters its protein structure upon binding proteasomal subunit S5a Proc. Natl. Acad. Sci. USA . 100, PubMedID: , DOI:
  71. Yee, C.S., et al. 2003. Generation of the R2 subunit of ribonucleotide reductase by intein chemistry: insertion of 3-nitrotyrosine at residue 356 as a probe of the radical initiation process Biochemistry. 42, PubMedID: 14661967, DOI:
  72. Cotton, G.J. and Muir, T.W. 2000. Generation of a dual-labeled fluorescence biosensor for Crk-II phosphorylation using solid-phase expressed protein ligation Chem. Biol. 4, PubMedID: 10780925, DOI:
  73. Grzybowska, B., Szweda, P. and Synowiecki, J. 2004. Cloning of the thermostable alpha-amylase gene from Pyrococcus woesei in Escherichia coli: isolation and some properties of the enzyme Mol. Biotechnol. . 26, PubMedID: 14764935, DOI:
  74. Romanelli, A., et al. 2004. Semisynthesis of a segmental isotopically labeled protein splicing precursor: NMR evidence for an unusual peptide bond at the N-extein-intein junction Proc. Natl. Acad. Sci. USA. 101, PubMedID: 15087498, DOI:
  75. Hemelaar, J.et al. 2004. Specific and covalent targeting of conjugating and deconjugating enzymes of ubiquitin-like proteins Mol. Cell Biol.. 24, PubMedID: 14673145, DOI:
  76. Takeda, S.,et al. 2004. Site-specific conjugation of oligonucleotides to the C-terminus of recombinant protein by expressed protein ligation Bioorg. Med. Chem. Lett.. 14(10), PubMedID: 15109622, DOI:
  77. Lue, R.Y., et al. 2004. Versatile protein biotinylation strategies for potential high-throughput proteomics J Am. Chem. Soc.. 126, PubMedID: 14746473, DOI:
  78. Durek, T., et al. 2004. Synthesis of fluorescently labeled mono- and diprenylated Rab7 GTPase J. Am. Chem. Soc.. 126, PubMedID: 15600338, DOI:
  79. Pezza, J.A., Allen, K.N. and Tolan, D.R. 2004. Intein-mediated purification of a recombinantly expressed peptide Chem Commun (Camb). 21, PubMedID: 15514791, DOI:
  80. Guo, C., et al. 2004. Intein-mediated fusion expression, high efficient refolding, and one-step purification of gelonin toxin Protein Expr Purif.. 37, PubMedID: , DOI:
  81. Durek T., Goody R.S and Alexandrov K. 2004. In vitro semisynthesis and applications of C-terminally modified Rab proteins Methods Mol Biol.. 283, PubMedID: 15197315, DOI:
  82. Kaiser, C.M., Chang, H.C., Agashe, V.R., Lakshmipathy, S.K., Etchells, S.A., Hayer-Hartl, M., Hartl, F.U. and Barral, J.M. 2006. Real-time observation of trigger factor function on translating ribosomes Nature. 444, PubMedID: 17051157, DOI:
  83. Kalia, J., et al. 2006. Reactivity of intein thioesters: appending a functional group to a protein Chembiochem.. 7, PubMedID: 16897799, DOI:
  84. Gottlieb, D., et al. 2006. Intein-mediated in vitro synthesis of lipidated Ras proteins Chem. Commun. (Camb). 21, PubMedID: 16391727, DOI:
  85. Wojtaszek, J., et al. 2006. LT CI, a novel chymotrypsin inhibitor of the potato I family from the earthworm Lumbricus terrestris. Purification, cDNA cloning, and expression Comp. Biochem. Physiol B. Biochem. Mol. Biol. . 143, PubMedID: , DOI:
  86. Demidov, V.V., and Broude, N.E. 2006. Profluorescent protein fragments for fast bimolecular fluorescence complementation in vitro Nat. Protoc.. 1, PubMedID: 17406301, DOI:
  87. Esipov, R.S., et. al. 2006. Production and purification of recombinant human glucagons overexpressed as intein fusion protein in Escherichia coli Protein Pept. Lett.. 13, PubMedID: 16712508, DOI:
  88. Grant, J., et al. 2006. The N terminus of GTP gamma S-activated transducin alpha-subunit interacts with the C terminus of the cGMP phosphodiesterase gammasubunit J. Biol. Chem. . 281, PubMedID: 16407279, DOI:
  89. Eckenroth, B., et al. 2006. Semisynthesis and characterization of mammalian thioredoxin reductase Biochem.. 45, PubMedID: 16618105, DOI:
  90. Lee, N.Y., et al. 2006. Structure and dynamics of the epidermal growth factor receptor C-terminal phosphorylation domain Protein Sci.. 15, PubMedID: 16597832, DOI:
  91. Shimizu, Y. and Ueda, T. 2006. SmpB triggers GTP hydrolysis of elongation factor Tu on ribosome by compensating for the lack of codon-anticodon interaction during trans-translation initiation J. Biol. Chem.. 281, PubMedID: 16601123, DOI:
  92. Muto, H., Nakatogawa, H., and Ito, K. 2006. Genetically Encoded but Nonpolypeptide Prolyl-tRNA Functions in the A Site for SecM-Mediated Ribosomal Stall Mol. Cell.. 22, PubMedID: 16713584, DOI:
  93. Villemagne, D., Jackson, R. and Douthwaite, J.A. 2006. Highly efficient ribosome display selection by use of purified components for in vitro translation J. Immunol Methods. 313, PubMedID: 16730021, DOI:
  94. Van Ooyen, A.J.J., Dekker, P., Huang, M., Olsthoorn, M.M.A., Jacobs, D.I., Colussi P.A., and Taron, C.H. 2006. Heterologous protein production in the yeast Kluyveromyces lactis FEMS Yeast Res.. 6, PubMedID: 16630278, DOI:
  95. Hackenberger, C. P., et. al. 2006. Expression of N-terminal Cys-protein fragments using an intein refolding strategy Bioorg. Med. Chem.. 14, PubMedID: , DOI:
  96. Sando, S., Kanatani, K., Sato, N., Matsumoto, H. Hohsaka, T. and Aoyama, Y. 2005. A small-molecule-based approach to sense codon-templated natural-unnatural hybrid peptides. Selective silencing and reassignment of the sense codon by orthogonal reacylation stalling at the single-codon level J. Am. Chem. Soc.. 127, PubMedID: 15926808, DOI:
  97. Colussi, P.A. and Taron, C.H. 2005. Kluyveromyces lactis LAC4 promoter variants that lack function in bacteria but retain full function in K. lactis Appl. Environ. Microbiol.. 71, PubMedID: 16269745, DOI:
  98. Shimizu, Y. Kanamori, T. and Ueda, T. 2005. Protein synthesus by pure translation systems Methods. 36, PubMedID: 16076456, DOI:
  99. Goodin, J. L., et al. 2005. Yersinia pestis outer membrane type III secretion protein YscC: expression, purification, characterization, and induction of specific antiserum Protein Expr. Purif. . 40, PubMedID: 15721783, DOI:
  100. Ge, X., et. al. 2005. Self-cleavable stimulus responsive tags for protein purification without chromatography J. Am. Chem. Soc. 127, PubMedID: 16089436, DOI:
  101. Girish, A., et al. 2005. Site-specific immobilization of proteins in a microarray using intein-mediated protein splicing Bioorg. Med. Chem. Lett.. 15, PubMedID: 15863295, DOI:
  102. Colussi, P.A., Specht, C.A. and Taron, C.H. 2005. Characterization of a nucleus-encoded chitinase from the yeast Kluyveromyces lactis Appl. Environ. Microbiol.. 71, PubMedID: 15932978, DOI:
  103. Ying, B.W., Taguchi, H., Kondo, M. and Ueda, T. 2005. Co-translational involvement of the chaperonin GroEL in the folding of newly translated polypeptides J. Biol. Chem.. 280, PubMedID: 15664980, DOI:
  104. Ingham, A.B., et al. 2005. A versatile system for the expression of nonmodified bacteriocins inEscherichia coli J. Appl. Microbiol. . 98, PubMedID: 15715871, DOI:
  105. Morassutti, C., et al. 2005. Expression of SMAP-29 cathelicidin-like peptide in bacterial cells by intein-mediated system Protein Expr. Purif.. 39, PubMedID: , DOI:
  106. Rebets, Y., et al. 2005. DNA -binding activity of LndI protein and temporal expression of the gene that upregulates landomycin E production in Streptomyces globisporus 1912 Microbiology . , PubMedID: , DOI:
  107. Kuruma, Y., Nishiyama, K., Shimizu, Y., Muller, M and Ueda, T. 2005. Development of a minimal cell-free translation system for the synthesis of presecretory and integral membrance proteins Biotechnol, Prog.. 21, PubMedID: 16080708, DOI:
  108. Theerthagiri, G., Eisenhardt, N., Schwarz, H. and Antonin, W. 2010. The nucleoporin Nup188 controls passage of membrane proteins across the nuclear pore complex  J. Cell Biol.  . 189, PubMedID: 20566687, DOI:
  109. Noto, T., Kurth, H., Kataoka, K., Aronica, L., DeSouza, L., Siu, K., Pearlman, R., Gorovsky, M. and Mochizuki, K. 2010. The tetrahymena argonaute-binding protein Giw1p directs a mature argonaute-siRNA complex to the nucleus Cell . 140 , PubMedID: 20211138, DOI:
  110. Shen, B.W., Heiter, D.F., Chan, S.H., Wang, H., Xu, S.Y., Morgan, R.D., Wilson, G.G. and Stoddard, B.L. 2010. Unusual target site disruption by the rare-cutting HNH restriction endonuclease PacI Structure . 18, PubMedID: 20541511, DOI:
  111. Lu, M., et. al. 2010. Purification of untagged HIV-1 reverse transcriptase by affinity chromatography Protein Expr. Purif.. 71, PubMedID: 20060474, DOI:
  112. Ueda, T., Kanamori, T., Ohashi, H. 2010. Ribosome display with the PURE technology Methods Mol Biol. . 607, PubMedID: 20204860, DOI:
  113. Chen, J., et al. 2010. Chemically ubiquitylated PCNA as a probe for eukaryotic translesion DNA synthesis Nat. Chem. Biol. . 6, PubMedID: 20208521, DOI:
  114. Feng, Z., et al. 2010. Codon optimization of the calf prochymosin gene and its expression in Kluyveromyces lactis World J. Microbiol. Biotechnol.. 26, PubMedID: , DOI:
  115. Collins, E. D., et. al. 2010. Cloning the human vitamin D receptor into the pTwin-1 expression vector J Steroid Biochem Mol Biol. , PubMedID: 20171280 , DOI:
  116. Karukurichi, K. et al. 2010. Analysis of p300/CBP histone acetyltransferase regulation using circular permutation and semisynthesis J. Am. Chem. Soc. . 132, PubMedID: 20063892, DOI:
  117. Yu, H. H., et. al. 2010. Expressed protein ligation for the preparation of fusion proteins with cell penetrating peptides for endotoxin removal and intracellular delivery Biochim Biophys Acta . , PubMedID: 20170629, DOI:
  118. Manconi, B., et. al. 2010. Expression, purification, phosphorylation and characterization of recombinant human statherin Protein Expr Purif.. 69, PubMedID: , DOI:
  119. Asahara, H. and Chong, S. 2010. In vitro genetic reconstruction of bacterial transcription initiation by coupled synthesis and detection of RNA polymerase holoenzyme Nucleic Acids Res. . 38, PubMedID: 20457746, DOI:
  120. Yuan W, et al. 2010. Gene synthesis of the bovine prochymosin gene and high-level expression in Kluyvermyces lactis Sheng Wu Gong Cheng Xue Bao. 26, PubMedID: 21141120, DOI:
  121. Rocha, S.N., et al. 2010. Heterologous expression of glucose oxidase in the yeastKluyveromyces marxianus Microb. Cell Fact.. 21, PubMedID: 20092622, DOI:
  122. Kuo, D., et al. 2010. Evolutionary divergence in the fungal response to fluconazole revealed by soft clustering Gen. Bio. . 11, PubMedID: 20653936, DOI:
  123. Lu, X., et al. 2010. Designed semisynthetic protein inhibitors of ub/ubl e1 activating enzymes J. Am. Chem. Soc. . 132, PubMedID: 20099854, DOI:
  124. Bhat, R. K. and Berger, S. 2007. New and easy strategy for cloning, expression, purification, and characterization of the 5S subunit of transcarboxylase from Propionibacterium f. shermanii Prep. Biochem. Biotechnol.. 37, PubMedID: 17134979, DOI:
  125. Murtas, G., Kuruma, Y., Bianchini, P., Diaspro, A., and Luisi, P.L. 2007. Protein synthesis in liposomes with a minimal set enzymes Biochem. Biophys. Res. Commun.  . 363, PubMedID: 17850764, DOI:
  126. Wamalwa, B.M., et al. 2007. High-Level Heterologous Expression of Bacillus halodurans Putative Xylanase Xyn11A (BH0899) in Kluyveromyces lactis Biosci. Biotechnol. Biochem.. 71, PubMedID: 17341817, DOI:
  127. Diao, H., et. al. 2007. Intein-mediated expression is an effective approach in the study of beta-defensins Biochem Biophys Res Commun. 357, PubMedID: 17445764, DOI:
  128. Paulick, M. G., et al. 2007. Synthetic analogues of glycosylphosphatidylinositol-anchored proteins and their behavior in supported lipid bilayers J. Am. Chem. Soc. . 129, PubMedID: 17715922, DOI:
  129. Kochinyan, S., et al. 2007. Use of intein-mediated phosphoprotein arrays to study substrate specificity of protein phosphatases Biotechniques. 42, PubMedID: 17269486 , DOI:
  130. Ohta, A., Murakami, H., Higashimura, E., and Suga, H. 2007. Synthesis of Polyester by Means of Genetic Code Reprogramming Chem. Biol. . 14, PubMedID: 18096500, DOI:
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Protein Expression and Purification: Selection Chart

Each NEB expression technology offers different advantages which enable you to choose the strategy that best suits your protein expression and purification needs.

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    Solutions for the Expression of Difficult Proteins

    NEB has a long history in recombinant protein expression and has developed a wide array of solutions for proteins that are difficult to express.