Expression and purification of functional recombinant CUL2•RBX1 from E. coli

Expression and purification of functional recombinant CUL2•RBX1 from E. coli
July 22, 2021 0 Comments

Cullin-2 (CUL2) primarily based cullin-RING ligases (CRL2s) comprise a household of ubiquitin E3 ligases that exist solely in multi-cellular organisms and are essential for mobile processes equivalent to embryogenesis and viral pathogenesis. CUL2 is the scaffold protein that binds one of the interchangeable substrate receptor modules, which consists of adaptor proteins and the substrate receptor protein. The VHL protein is a substrate receptor identified to focus on hypoxia-inducible issue α (HIF1α) for ubiquitination and degradation. Because of its essential function within the ubiquitination of necessary mobile components equivalent to HIF1α, CRL2s have been investigated for his or her organic features and the event of novel therapeutics towards illnesses. Given the significance of CRL2s in organic and biomedical analysis, strategies that effectively produce functional CUL2 proteins will significantly facilitate research on the mechanism and regulation of CRL2s. Here, we report two cost-effective programs for the expression and purification of recombinant human CUL2 from E. coli cells. The purified CUL2

proteins have been ~ 95% pure, may bind their substrate receptor modules, and have been enzymatically read more

Bladder Cancer QC

June 21, 2021 0 Comments

The microbiota isolated from the urine of patients with bladder carcinoma shows a significantly higher compositional abundance of some bacterial genera compared to the urine of healthy patients. Our objective was to compare the microbiota composition of cancerous tissues and urine samples collected from the same group of patients to improve the precision of diagnostic measures.

Tissue samples were collected from patients during the removal of cancerous tissue by transurethral resection. In parallel, urine samples were obtained by means of a transurethral resectoscope from the same patients. The V3-V4 region of the bacterial 16S rRNA gene was sequenced and analyzed using the Kraken pipeline. In the case of four patients, the duplicate microbiota analysis of distant parts of the cancerous tissues was highly reproducible and independent of the tissue collection site from any given patient. Akkermansia, Bacteroides, Clostridium sensu stricto, Enterobacter, and Klebsiella, as “five suspect genera”, were overrepresented in tissue samples compared to urine. read more

Xpert Carba-R QC Panel M219

June 17, 2021 0 Comments

The Xpert Carba-R QC Panel M219 is designed for in vitro use as quality control to monitor the detection and differentiation of 5 beta-lactamase gene sequences (blaKPC, blaNDM, blaVIM, blaOXA-48, and blaIMP-1) when tested with the Cepheid Xpert Carba-R Assay in the GeneXpert instrument.

Gene sequences are associated with non-susceptibility to carbapenems. in gram-negative bacteria. The worldwide spread of bacteria not susceptible to carbapenems is a critical public health problem. 1,2 These bacteria are often resistant to all beta-lactam agents and are often co-resistant to multiple classes of other antimicrobial agents, leaving very few treatment options. read more

The effects of LH inhibition with cetrorelix on cumulus cell gene expression during the luteal phase under ovarian coasting stimulation in cattle

April 24, 2020 0 Comments

Cumulus cells have an important role to play in the final preparation of the oocyte before ovulation. During the final phase of follicular differentiation, FSH levels are low and LH maintains follicular growth; however, it is not known if at that time LH has an influence on cumulus cells inside the follicle. In humans, LH is often inhibited to avoid a premature ovulatory LH surge. This procedure provides a tool to investigate the role of LH in follicular development. In this study, we investigated the impact of suppressing LH using the GnRH antagonist cetrorelix during an ovarian coasting stimulation protocol on the transcriptome of bovine cumulus cells (CC). Oocytes were collected twice from 6 dairy cows. For the first collection, the cows received FSH twice daily for 3 d, followed by FSH withdrawal for 68 h as a control protocol. For the second collection, the same stimulation protocol was used, but the cows also received, starting on day 2 of FSH stimulation, a GnRH antagonist once a day until recovery of the cumulus-oocyte complexes (COC). Half of the COC were subjected to in vitro maturation, fertilization, and culture to assess blastocyst rates. The other half of the COC underwent microarray analysis (n = 3 cows, 2 treatments, 6 oocyte collections) and qRT-PCR (n = 6 cows: 3 microarray cows +3 other cows, 2 treatments, 12 oocyte collections). The differential expression of specific genes was confirmed by RT-qPCR: decrease of ATP6AP2, SC4MOL, and OSTC and increase of PTGDS in the LH-inhibited condition. The global transcriptomic analysis of cumulus cells demonstrated that the inhibition of LH secretion may decrease survival and growth of the follicle. Moreover, the results suggested that LH may be important to cumulus for the maintenance of cellular mechanisms such as global RNA expression, protein and nucleic acid metabolism, and energy production. These results support the hypothesis that LH support is important during the final part of follicle maturation through its influence on the cumulus cells.