Total protein concentrations in cell lysates were determined using a Solarbio BCA kit. Cell lysates were separated on 5–15% SDS-PAGE gels and transferred onto PVDF membranes (Merck Millipore) that were then blocked with 5% non-fat milk in TBST (0.05% Tween-20 in Tris-buffered Saline) before...
fumC(fumarase C),yjbJ(putative stress-response protein),hchA(protein/nucleic acid deglycase 1),frdD(fumarate reductase subunit D),elaB(tail-anchored inner membrane protein) andrraB(ribonuclease E inhibitor protein B) genes were cloned in a low-copy plasmid (pMMB) under the control of an ...
The remaining precipitate was washed with cold acetone for three times, and total protein was quantified by BCA kit. After trypsin digestion, peptide was desalted by Strata X C18 SPE column (Phenomenex) and vacuum dried. Peptide was reconstituted in 0.5 M triethylamine bicarbonate and mixed with...
Total protein concentration of plasma was measured by BC Assay (Protein assay kit, Uptima, Montlucon, France). 2.6. RNA isolation, cDNA synthesis and gene expression analyses Total RNA was isolated from cultured cells using the RNeasy Mini kit (Qiagen, Hilden, Germany). RNA was reversely ...
This leads to localised cellular stress and subsequent expression of antioxidant protein-encoding genes including peroxiredoxin 1. This hypothesised functional explanation is consistent with the histological examination of the 'lactating' crop presented here and in other studies....
RNA isolation, cDNA synthesis and RT-PCR For global gene expression analyses, the total RNA was purified in triplicate from isolated spermatocytes and hepatocytes using RNeasy® Mini Kit spin columns (Qiagen, Valencia, CA, USA). Validation of the RNA quality was performed using the RNA 6000 ...
The gene coding ribosomal protein L5 (54775, RibL5) was used as an internal control according to the references [8, 53, 54] and the analysis of our transcriptomic data. For single-strand cDNA synthesis, the PrimeScript RT regent Kit with genomic DNA Eraser (TaKaRa) was used to perform ...
Mouse mammary tumor virus (MMTV) is a betaretrovirus that causes breast cancer in mice. The mouse mammary epithelial cells are the most permissive cells for MMTV, expressing the highest levels of virus upon infection and being the ones later transformed
More highly expressed genes are involved in protein synthesis and oxidative phosphorylation, which is consistent with the high rates of fungal growth in the culture conditions. Approximately 99% of the NMP codifying genes are expressed. RNAseq also aids in the identification of new genes. Based on...
(5–7 days after inoculation) with few remainingT. gayanumliving cells and rich in aphelid plasmodia, resting spores and zoospores. To minimize algal overrepresentation, we maintained the cultures in the dark. Total RNA was extracted with the RNeasy mini Kit (Qiagen), quantified by Qubit (...