Expression, Purification, and Antiserum Production of the Truncated UL31 Protein of Herpes Simplex Virus 1

Zou, Xiaofang; Zuo, Xu; Wang, Yuanfang; Ou, Xiaowen; Li, Yiwen; Liu, Delong; Gan, Weidong; Lu, Manjiao; Chen, Qiusan; Peng, Hao; Hong, Gengde; JinYu, Lin; Li, Meili; Cai, Mingsheng

doi:10.21859/ijb.1609

Cited by 4 publications

(2 citation statements)

References 39 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The intermediate temperature of 25 °C was best suited for our experimental work (Figure 3). The IPTG induction of recombinant proteins like human RNase L, human growth hormone (hGH), and several other proteins at lower temperatures in bacteria favors relatively higher yield of the expressed protein in the soluble fraction of the cell [41][42][43][44][45][46][47]. The experimental conditions optimized above for the recombinant SigB expression provided a high…”

Section: Discussionmentioning

confidence: 99%

Expression, Purification, and In Silico Characterization of Mycobacterium smegmatis Alternative Sigma Factor SigB

et al. 2022

View full text Add to dashboard Cite

Sigma factor B (SigB), an alternative sigma factor (ASF), is very similar to primary sigma factor SigA (σ70) but dispensable for growth in both Mycobacterium smegmatis (Msmeg) and Mycobacterium tuberculosis (Mtb). It is involved in general stress responses including heat, oxidative, surface, starvation stress, and macrophage infections. Despite having an extremely short half-life, SigB tends to operate downstream of at least three stress-responsive extra cytoplasmic function (ECF) sigma factors (SigH, SigE, SigL) and SigF involved in multiple signaling pathways. There is very little information available regarding the regulation of SigB sigma factor and its interacting protein partners. Hence, we cloned the SigB gene into pET28a vector and optimized its expression in three different strains of E. coli, viz., (BL21 (DE3), C41 (DE3), and CodonPlus (DE3)). We also optimized several other parameters for the expression of recombinant SigB including IPTG concentration, temperature, and time duration. We achieved the maximum expression of SigB at 25°C in the soluble fraction of the cell which was purified by affinity chromatography using Ni-NTA and further confirmed by Western blotting. Further, structural characterization demonstrates the instability of SigB in comparison to SigA that is carried out using homology modeling and structure function relationship. We have done protein-protein docking of RNA polymerase (RNAP) of Msmeg and SigB. This effort provides a platform for pulldown assay, structural, and other studies with the recombinant protein to deduce the SigB interacting proteins, which might pave the way to study its signaling networks along with its regulation.

show abstract

Section: Discussionmentioning

confidence: 99%

Expression, Purification, and In Silico Characterization of Mycobacterium smegmatis Alternative Sigma Factor SigB

et al. 2022

View full text Add to dashboard Cite

show abstract

“…In these regards immunogenicity of the multi-epitopic recombinant glycoproteins virus: implications may be concerned (48). There are also several biotechnological research on the virus (49)(50)(51)(52)(53)(54)(55)(56)(57)(58)(59)(60)(61), each of which can be helpful in applying the results. There are several limitations to this study.…”

Section: Discussionmentioning

confidence: 99%

Importance of miR-125a-5p and miR-122-5p expression in patients with HBV infection

Lak

Yaghobi

Garshasbi

2020

Cell Mol Biol (Noisy-le-grand)

View full text Add to dashboard Cite

MicroRNAs (miRNAs) as a small RNA and post-transcriptional modulator are shown to have regulatory effects for different cellular activities and pathways, such as metabolism, virus replication and also cell growth. In addition, miRNAs can regulate the replication of the hepatitis B virus (HBV). Therefore, the expression profile of miRNAs was evaluated in HBV infected patient groups and healthy controls. The expression levels of following microRNAs (as noninvasive biomarkers) were compared in two experimental (those with various stages of HBV infection) and control groups to evaluate their diagnosis ability: miR-122-5p, miR-125a-5p, miR-199a-3p, miR-210-5p, miR-205-5p, miR-155-5p, miR-372-5p, and miR-1-5p. RNA extraction was performed for 45 serum samples. The miRCURY LNA™ Universal RT-miRNA-PCR system and miRNA PCR panels were used for measuring microRNA expression profiles. To normalize quantitative values, the endogenous reference by UniSp6 expression was used.Serum miR-125a-5p and miR-122-5p were significantly higher in patients in different stages of HBV infection (p<0.001) than in controls (p<0.001). Receiver operating characteristic (ROC) curve analyses suggested that serum miR-125a-5p and miR-122-5p have significant diagnostic value for HBV infection. A significant difference was not found in terms of serum levels of other miRs (miR-199a-3p, miR-210-5p, miR-205-5p, miR-155-5p, miR-372-5p and miR-1-5p). Results suggest that miR-125a-5p and miR-122-5p can be used as possible noninvasive biomarkers for monitoring of HBV infection need to confirm in future completed studies.

show abstract

The Epstein–Barr virus small capsid protein BFRF3 disrupts the NF‐кB signaling pathway by inhibiting p65 activity

Li,

Liu

et al. 2024

The FASEB Journal

View full text Add to dashboard Cite

Epstein–Barr virus (EBV), a common gamma herpesvirus, establishes a life‐long latent infection in the host to defend against innate immune recognition, which is closely related to a variety of malignant tumors, but its specific mechanism is unclear. BFRF3, an EBV‐encoded small capsid protein, is mainly involved in the assembly of the viral capsid structure and the maintenance of its stability. Here, we showed that BFRF3 can inhibit TNF‐α‐mediated NF‐кB promoter activation. Moreover, BFRF3 downregulates NF‐кB‐mediated promoter activation and transcription of inflammatory cytokines, including IL‐6 and IL‐8. Dual‐luciferase reporter assay demonstrated that BFRF3 restrains NF‐кB promoter activity at or below the p65 level, and coimmunoprecipitation analysis revealed that BFRF3 not only interacts with p65 but also binds to its critical truncated Rel homology domain (RHD) and transcriptional activation domain (TAD). However, BFRF3 does not affect the dimerization of p65‐p50, but overexpression of BFRF3 reduces the nuclear accumulation of p65, and the phosphorylation of p65 (Ser536) is repressed during BFRF3 transfection and EBV lytic infection, which promotes the proliferation of EBV. Overall, our study suggested that BFRF3 may play a crucial role in antiviral immunity to defend against EBV infection by inhibiting NF‐κB activity.

show abstract

Expression, Purification, and Antiserum Production of the Truncated UL31 Protein of Herpes Simplex Virus 1

Cited by 4 publications

References 39 publications

Expression, Purification, and In Silico Characterization of Mycobacterium smegmatis Alternative Sigma Factor SigB

Expression, Purification, and In Silico Characterization of Mycobacterium smegmatis Alternative Sigma Factor SigB

Importance of miR-125a-5p and miR-122-5p expression in patients with HBV infection

The Epstein–Barr virus small capsid protein BFRF3 disrupts the NF‐кB signaling pathway by inhibiting p65 activity

Contact Info

Product

Resources

About