Protein expression and purification最新文献

{"title":"Mini review for niche downstream processes","authors":"Tsutomu Arakawa ,&nbsp;Daisuke Ejima ,&nbsp;Yui Tomioka ,&nbsp;Chiaki Sakuma ,&nbsp;Teruo Akuta","doi":"10.1016/j.pep.2025.106690","DOIUrl":"10.1016/j.pep.2025.106690","url":null,"abstract":"<div><div>We review here several niche downstream purification processes that are not covered by other articles in this special issue. The first is the use of activated carbon to capture contaminants and clarify culture medium for purification of proteins, including antibodies. Flow-through operation of the activated carbon filter showed over 80 % recovery of antibodies with 10--fold reduction of host cell proteins and effective lipopolysaccharide and virus removal. The second is salt or arginine-tolerant column chromatography, in which the respective resins, such as hydroxyapatite and mixed-mode resins, can bind proteins in the presence of salt or arginine at high concentrations. Effective use of arginine was also suggested in size exclusion and affinity chromatography. The last is the isolation of proteins using gel electrophoresis and simple extraction procedure. These technologies offer simple and cost-effective methods for purifying proteins and protein complexes in the native state.</div></div>","PeriodicalId":20757,"journal":{"name":"Protein expression and purification","volume":"230 ","pages":"Article 106690"},"PeriodicalIF":1.4,"publicationDate":"2025-02-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143465114","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Expression and purification of recombinant glutaredoxin 1 and protection against oxidative stress injury during cerebral ischemia-reperfusion injury","authors":"Zi-teng Li,&nbsp;Tong Lin,&nbsp;Yu Sun,&nbsp;Xin-yi Wang,&nbsp;Yi-xuan Yang,&nbsp;Li Gan,&nbsp;Jia-ming Xu,&nbsp;Xu-ting Wei,&nbsp;Huang-qing Zhu,&nbsp;Wei-chun Zhao,&nbsp;Zhen-hong Zhu","doi":"10.1016/j.pep.2025.106689","DOIUrl":"10.1016/j.pep.2025.106689","url":null,"abstract":"<div><div>Glutaredoxin (Grx) is a small molecular protein widely found in both prokaryotes and eukaryotes, serving various biological functions, including participation in redox reactions and exerting anti-apoptotic effects[1]. To evaluate the protective effect of recombinant Grx1 against oxidative stress, we constructed the pET-30a (+)/Grx1 recombinant plasmid and performed soluble expression and purification of the recombinant Grx1. In vitro experiments, including ABTS and DPPH radical scavenging assays, showed that recombinant Grx1 has significant antioxidant activity. Reactive oxygen species detection revealed that the levels of reactive oxygen species in the Grx1 treatment group decreased by 33.01 % compared to the H₂O₂ group. Flow cytometry analyses indicated that the number of apoptotic cells in the Grx1 treatment group decreased by 23.51 % relative to the H₂O₂ group. Additionally, qRT-PCR analysis showed that Grx1 significantly reduced the expression levels of genes such as IL-1β, TNF-α, IL-6, and caspase-3 in PC12 cells. <em>In vivo</em>, recombinant Grx1 was utilized to treat cerebral ischemia-reperfusion injury (CIRI). Histological staining revealed that recombinant Grx1 significantly mitigated hippocampal tissue damage. Western blotting analysis demonstrated that Grx1 can reduce neuronal apoptosis following CIRI by decreasing Bax expression while increasing Bcl-2 expression. Furthermore, Grx1 was shown to modulate the HO-1/Nrf2 signaling pathway by elevating the expression of Nrf2 and HO-1. In summary, this study successfully overexpressed biologically active Grx1 in <em>E</em>. <em>coli,</em> and confirms that recombinant Grx1 exhibits remarkable antioxidant activity in both <em>in vitro</em> and <em>in vivo</em> experiments, effectively alleviating oxidative stress damage associated with ischemic stroke.</div></div>","PeriodicalId":20757,"journal":{"name":"Protein expression and purification","volume":"230 ","pages":"Article 106689"},"PeriodicalIF":1.4,"publicationDate":"2025-02-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143459212","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"StxB fusion strategy for immunogenic enhancement: Recombinant expression of AcAMP antimicrobial peptide","authors":"Ehsan Zamani ,&nbsp;Jamil Zargan ,&nbsp;Hossein Honari ,&nbsp;Hani Keshavarz Alikhani","doi":"10.1016/j.pep.2025.106688","DOIUrl":"10.1016/j.pep.2025.106688","url":null,"abstract":"<div><h3>Background and objectives</h3><div>Polyclonal antibodies are essential for identifying compounds like peptides. However, many peptides exhibit low immunogenicity, resulting in reduced antibody yields. To address this, fusing peptides with immunogenic fusion proteins has been proposed. This study aimed to enhance the immunogenicity of the AcAMP peptide by fusing it with StxB as a fusion protein, producing the recombinant construct in <em>Escherichia coli,</em> and comparing its antibody titration to recombinant AcAMP in mice.</div></div><div><h3>Methods</h3><div>The <em>acamp</em> gene, containing BamHI and SalI restriction sites, was amplified from the pUC57 plasmid via PCR and cloned into the pET28a (+)-StxB expression vector. The pET28a (+)-AcAMP construct was prepared as previously described. Both constructs were expressed in <em>E. coli</em> BL21 (DE3) cells, induced with IPTG, and purified using nickel affinity chromatography. Recombinant proteins were confirmed by SDS-PAGE and Western blotting. Mice were immunized with purified proteins, and serum IgG titration was assessed using indirect ELISA.</div></div><div><h3>Results</h3><div>PCR amplification and enzymatic digestion verified the successful construction of the pET28a (+)-<em>StxB-acamp</em> vector. SDS-PAGE and Western blotting identified recombinant AcAMP and AcAMP-StxB proteins at 9 and 17 kDa, respectively. ELISA revealed significantly higher antibody titers for the recombinant AcAMP-StxB protein compared to recombinant AcAMP.</div></div><div><h3>Conclusion</h3><div>Fusing the AcAMP peptide with the StxB protein enhanced immunogenicity and increased antibody production. This approach may improve expression conditions and immunogenicity for peptides of this family, advancing their use in identification studies.</div></div>","PeriodicalId":20757,"journal":{"name":"Protein expression and purification","volume":"229 ","pages":"Article 106688"},"PeriodicalIF":1.4,"publicationDate":"2025-02-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143429167","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Production of recombinant coiled coil silk proteins for materials synthesis","authors":"Patrick J. Shilling ,&nbsp;Luisa Pontes-Braz ,&nbsp;Lachlan Mitchell ,&nbsp;Linda Howell ,&nbsp;Prem Veneer ,&nbsp;Srinivasan Jayashree ,&nbsp;Laura A. Castelli ,&nbsp;Tam Pham ,&nbsp;Louis Lu ,&nbsp;Bei Wang ,&nbsp;K.Y Benjamin Yeo ,&nbsp;Surekha Nimma ,&nbsp;Lyndall Briggs ,&nbsp;Caitlin Johnston ,&nbsp;Michelle Michie ,&nbsp;Tara D. Sutherland","doi":"10.1016/j.pep.2025.106683","DOIUrl":"10.1016/j.pep.2025.106683","url":null,"abstract":"<div><div>Rational design of fundamentally new advanced materials would be facilitated by availability of polymers with controlled monomer sequence. Recombinant proteins offer polymers with controlled monomer sequence but are underrepresented in material science, in part because suitable proteins cannot be produced at commercial levels in recombinant systems. The silk proteins of honeybees fulfil the requirements for rational materials design and can be produced at commercially viable levels. In this study we compare recombinant expression of these silks in bacteria, yeast and insect cells to identify the most suitable method of silk protein production. Yeast and insect cell lines are unlikely to be suitable expression platforms for these silks as the recombinant proteins were degraded, expression levels were low or absent, and host cell protein levels were high. We confirm that expression into <em>E. coli</em> inclusion bodies using defined media offers high level expression and to date is the best expression system for these proteins.</div></div>","PeriodicalId":20757,"journal":{"name":"Protein expression and purification","volume":"229 ","pages":"Article 106683"},"PeriodicalIF":1.4,"publicationDate":"2025-02-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143374397","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"High-affinity nanobodies targeting IL-12B for the detection of fluorescence resonance energy transfer","authors":"Jing Hu ,&nbsp;Wenxuan Feng ,&nbsp;Jianchuan Wen ,&nbsp;Siyu Zhou ,&nbsp;Zengchao Sun ,&nbsp;Shuaiying Zhao ,&nbsp;Shaojue Guo ,&nbsp;Hui Wang ,&nbsp;Yong Geng","doi":"10.1016/j.pep.2025.106681","DOIUrl":"10.1016/j.pep.2025.106681","url":null,"abstract":"<div><h3>Aims</h3><div>IL-12B, a subunit of the IL-23 family of cytokines, plays a crucial role in various diseases such as viral infections, autoimmune disorders, and tumors. This study aimed to identify high-affinity nanobodies that bind to distinct epitopes of IL-12B to and assess their potential for therapeutic and diagnostic applications, particularly through fluorescence resonance energy transfer(FRET) to evaluate their ability to target IL-12B.</div></div><div><h3>Methods</h3><div>IL-12B protein was expressed in eukaryotic cells and used to immunize camels to induce an immune response. Camel-derived anti-IL-12B nanobodies were isolated and screened via phage display to identify those with high specificity and affinity for IL-12B. Binding affinity and epitope interactions were further analyzed using high-performance liquid chromatography (HPLC) and ForteBio Octet assays. A FRET-based assay was developed to evaluate protein interactions for precise therapeutic targeting.</div></div><div><h3>Results</h3><div>Several high-affinity nanobodies targeting IL-12B were successfully generated. These nanobodies exhibited strong binding to various epitopes of IL-12B. Screening by HPLC and ForteBio Octet confirmed their high specificity and affinity, while fluorescence analysis demonstrated efficient energy transfer between thenanobodies, indicating successful interactions.</div></div><div><h3>Conclusions</h3><div>This study identified high-affinity nanobodies against IL-12B and used FRET to characterize their interactions. These nanobodies show promise for therapeutic potential targeting IL-12B-related diseases, including viral infections, autoimmune disorders, and cancer. However, further clinical studies are needed to fully explore their potential for diagnostic and therapeutic applications.</div></div>","PeriodicalId":20757,"journal":{"name":"Protein expression and purification","volume":"229 ","pages":"Article 106681"},"PeriodicalIF":1.4,"publicationDate":"2025-02-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143374386","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Expression and biochemical characterization of a novel NAD+-dependent xylitol dehydrogenase from the plant endophytic fungus Trichoderma gamsii","authors":"Shuping Fei ,&nbsp;Wenxiu Hu ,&nbsp;Jingwen Shu ,&nbsp;Ruirui Zhao ,&nbsp;Jiatong Zhao ,&nbsp;Mengwei Jiang ,&nbsp;Wenwen Wu ,&nbsp;Chaoqun Lian ,&nbsp;Wanggang Tang","doi":"10.1016/j.pep.2025.106687","DOIUrl":"10.1016/j.pep.2025.106687","url":null,"abstract":"<div><div>Xylitol dehydrogenase (XDH; EC 1.1.1.9), encoded by the <em>XYL2</em> gene, is a key enzyme in the fungal xylose metabolic pathway. In this work, a putative XDH from the plant endophytic fungus <em>Trichoderma gamsii</em> (TgXDH) was hetero-expressed in <em>Escherichia coli</em> BL21(DE3), purified to the homogeneity, and biochemically characterized. Sequence analysis revealed that TgXDH is 363 amino acids long and belongs to the zinc-containing medium-chain alcohol dehydrogenase superfamily. The size-exclusion chromatography analysis and SDS-PAGE showed that the purified recombinant TgXDH had a native molecular mass of ∼155 kDa and was composed of four identical subunits of molecular mass of ∼39 kDa. The optimum temperature and pH of this enzyme were 25 °C and pH 9.5, respectively. Kinetic analysis showed that it is an NAD⁺-dependent enzyme that has a polyol substrate preference (based on <em>k</em>_cat/<em>K</em>_m) in the order xylitol &gt; ribitol ≈ <span>d</span>-sorbitol. The <em>K</em>_m values for NAD⁺ with these three polyols ranged from 0.23 to 0.70 mM. Moreover, TgXDH showed high substrate affinities as compared to most of its homologs. The <em>K</em>_m values for xylitol, ribitol, and <span>d</span>-sorbitol were 5.23 ± 0.68 mM, 8.01 ± 1.22 mM, and 12.34 ± 1.37 mM, respectively. Collectively, the results will contribute to understanding the biochemical properties of a novel XDH from the filamentous fungi and provide a promising XDH for industrial production of ethanol.</div></div>","PeriodicalId":20757,"journal":{"name":"Protein expression and purification","volume":"229 ","pages":"Article 106687"},"PeriodicalIF":1.4,"publicationDate":"2025-02-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143365722","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Optimization on cell lysis and capture process of human adenovirus type 5 produced in suspension HEK293 cells","authors":"Sha Yi,&nbsp;Xiaoxu Gu,&nbsp;Youping Jin,&nbsp;Fei Wang,&nbsp;Junjun Jiang","doi":"10.1016/j.pep.2025.106686","DOIUrl":"10.1016/j.pep.2025.106686","url":null,"abstract":"<div><div>Adenovirus (Adv) is increasingly recognized for its significance in the fields of gene therapy and viral vector vaccines. The diverse applications in clinical trials and fundamental research necessitate the development of environmentally and economically sustainable purification processes that are straightforward and scalable for both academic and industrial contexts. In the initial segment of this study, we evaluated the lysis efficiency of polysorbate 20 (PS20) in comparison to polysorbate 80 (PS80). Our findings indicated that the viability HEK293 could be reduced to approximately 13 %, with a detectable Adv5 concentration of average 1.62 × 10⁹ ifu/mL in the supernatant after an incubation in 1.0 % (w/w) PS20 or PS80 buffer. In the subsequent portion of this research, we employed a high-throughput static binding capacity (SBC) screening tool in conjunction with on-column dynamic binding capacity (DBC) validation to concurrently assess the binding capacity of Adv5 on nine different types of anion exchange media. The results demonstrated that both Sartobind Q membrane and POROS 50HQ resin exhibited binding capacities exceeding 1.0 × 10¹³ vp/mL under the testing conditions.</div></div>","PeriodicalId":20757,"journal":{"name":"Protein expression and purification","volume":"229 ","pages":"Article 106686"},"PeriodicalIF":1.4,"publicationDate":"2025-02-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143348494","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Cloning, expression, and characterization of collagen galactosyltransferases from human, sponge, and sea walnut","authors":"Jeong Seon Kim,&nbsp;Tingfei Chen,&nbsp;Botao Zhang,&nbsp;Tristin M. Miller,&nbsp;Marisa E. Gilliam,&nbsp;Houfu Guo","doi":"10.1016/j.pep.2025.106685","DOIUrl":"10.1016/j.pep.2025.106685","url":null,"abstract":"<div><div>Collagen is an extracellular matrix protein conserved across animals and viruses, with its function regulated by post-translational modifications of lysine residues. Specifically, certain lysine residues in collagen are hydroxylated to form hydroxylysine, which serves as an attachment site for hydroxylysine-linked glycosylation. This glycosylation process is initiated by collagen galactosyltransferases from the GT25 family, also known as GLT25D or COLGALT proteins. Despite their biological importance, efficient methods for expressing and isolating GLT25Ds have yet to be fully developed, and the biochemical mechanisms underlying their function still need to be better understood. To address this, we performed sequence alignment and phylogenetic analyses of GLT25Ds across vertebrates, invertebrates, and viruses. Using sponge (<em>amphimedon queenslandica</em>) GLT25D as a model, we established a bacterial expression, purification, and assay protocol. Sponge GLT25D expressed robustly in <em>E. coli</em> strain BL21 and demonstrated enzymatic activity comparable to human GLT25D1 from mammalian cells. Kinetic parameters and the effects of time, temperature and pH on enzymatic activity were characterized for both enzymes. AlphaFold structural modeling and sequence alignment revealed an EXD motif and a conserved leucine in a pocket of the second Rossmann-fold domain of sponge GLT25D, suggesting this pocket as the active site. Using the standardized bacterial expression, purification, and assay protocol, we screened GLT25Ds from various vertebrate and invertebrate species. Notably, the sea walnut (<em>mnemiopsis leidyi</em>) GLT25D exhibited superior expression levels and robust enzymatic activity. This established method provides a strong foundation for future bioengineering efforts, structure-function analyses, and the development of GLT25D inhibitors.</div></div>","PeriodicalId":20757,"journal":{"name":"Protein expression and purification","volume":"229 ","pages":"Article 106685"},"PeriodicalIF":1.4,"publicationDate":"2025-02-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143190255","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Differentially labeled flaviviral protease-cofactor complex for NMR spectroscopic applications","authors":"Ajith Kumar","doi":"10.1016/j.pep.2025.106684","DOIUrl":"10.1016/j.pep.2025.106684","url":null,"abstract":"<div><div>Flaviviruses such as Dengue, Zika and West-Nile viruses have a positive strand RNA genome which is translated to a polyprotein inside the host cell. The viral polypeptide is matured to its constituents by the enzymatic action of NS2B-NS3 protease-cofactor complex. The flaviviral protease-cofactor complex attracted a lot of interest recently because of its potential for therapeutic intervention and the unique nature of catalysis where the peptide cofactor regulates the enzymatic activity. Obtaining the enzyme and cofactor differentially labeled with naturally abundant nuclei and NMR active nuclei respectively will be helpful in reducing the spectral complexity by making the enzyme invisible in a multidimensional NMR spectrum while only showing peaks from the cofactor. This will enable one to study the properties of the cofactor in isolation using NMR spectroscopy. Here, I have used a strategy for selectively labeling the cofactor within the complex with NMR active nuclei while peaks from the enzyme were rendered invisible. The protocol used here takes advantage of an ‘on-column unfolding’ step during the initial Ni-NTA chromatography to separate the enzyme and cofactor in unfolded conditions. The labeled cofactor was then allowed to fold in the presence of an unlabeled enzyme to obtain a differently labeled complex. We compared the ¹H-¹⁵N HSQC spectrum of the differently labeled, wild type and free cofactor to ensure that the cofactor attained the desired fold within the complex. The protocol is scalable, inexpensive and can be applied to other two-component enzyme systems where a peptide cofactor is essential for the folding of an enzyme.</div></div>","PeriodicalId":20757,"journal":{"name":"Protein expression and purification","volume":"229 ","pages":"Article 106684"},"PeriodicalIF":1.4,"publicationDate":"2025-02-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143103838","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Purification and characterization of the intrinsically disordered Arabidopsis thaliana protein SOG1","authors":"Kim Mignon ,&nbsp;Margot Galle ,&nbsp;Rani Van der Eecken ,&nbsp;Sarah Haesaerts ,&nbsp;Manon Demulder ,&nbsp;Henri De Greve ,&nbsp;Lieven De Veylder ,&nbsp;Remy Loris","doi":"10.1016/j.pep.2025.106678","DOIUrl":"10.1016/j.pep.2025.106678","url":null,"abstract":"<div><div>SOG1, a transcription factor consisting of a folded NAC (NAM-ATAF-CUC2) domain and an intrinsically disordered C-terminal domain (CTD), co-ordinates the DNA damage response in plants. Here we compare different methods to express and purify recombinant full length <em>Arabidopsis thaliana</em> SOG1. Expression in Sf9 insect cells results in a protein that contains a phosphorylated site that is possibly located on the T423 site in the CTD. This site is reported to be phosphorylated <em>in planta</em> upon aluminium toxicity stress and may affect the transcriptional activity of SOG1 in an yet undetermined way. Expression of SOG1 in <em>E. coli</em> BL21 (DE3) leads to the formation of inclusion bodies, a problem that is resolved by using a cleavable SUMO solubility tag. The resulting protein is not phosphorylated and represents the transcriptional inactive state of SOG1. Both protein preparations show similar CD spectra and melting temperatures. SEC-MALS determined that the proteins, like other NAC transcription factors, form a dimer in solution. Both proteins are also highly non-globular as determined by analytical SEC and are likely stretched out due to their disordered CTD. In electromobility shift assays, both insect and <em>E. coli</em> purified SOG1 proteins bind to a DNA fragment from the promoter region of <em>SMR5</em>, a well established target gene of SOG1, showing the functionality of both purified proteins.</div></div>","PeriodicalId":20757,"journal":{"name":"Protein expression and purification","volume":"229 ","pages":"Article 106678"},"PeriodicalIF":1.4,"publicationDate":"2025-01-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143075110","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}