A new device-mediated miniprep method

Baryshev, Mikhail; Dmitrijs, Merkulov; Ivan, Mironov

doi:10.1186/s13568-022-01360-7

Cited by 2 publications

(1 citation statement)

References 27 publications

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“…The pET-21a (+)-REJ was isolated with the ZymoPURE™ II Plasmid Maxiprep kit according to the manufacturer's protocol. [16] After extraction and purification, the pET-21a (+)-REJ was sent to the King Fahd medical research center, Jeddah, KSA, for sequencing to prove the identity of the plasmid sequence. The resulting pET-21a (+)-REJ sequence was analyzed using the Thermo Hitachi ABI Applied Biosystems 3500XL Genetic DNA Analyzer, Japan.…”

Section: Plasmid Dna Purification and Extractionmentioning

confidence: 99%

Cloning and Expression of Receptor of Egg Jelly Protein of Polycystic Kidney Disease 1 Gene in Human Receptor of Egg Jelly Protein

Sonbol¹,

AlRashidi²

2022

Pharmacophore

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Autosomal dominant polycystic kidney disease (ADPKD) is caused by a mutation in the polycystic kidney disease1 (PKD1) gene, which is responsible for 85% of ADPKD cases. The PKD1 gene encodes a polycystin-1 (PC1) protein that has a large extracellular area containing many polypeptide motifs. The extracellular region of PC1 includes several well-defined peptide domains that show that it has been involved in cell-cell and/or cell-matrix interactions. One of the regions that we focused on in this study is the receptor of the egg jelly (REJ) domain. This study conveys novel findings, in which we utilized a successful methodology to clone and express the REJ protein. The REJ gene is located in many exons separated by introns, which made it impossible to clone the whole REJ region. Therefore, synthetic DNA technology was used to clone a fragment located on exon 15 of the REJ gene. The PCR technology was used to amplify the REJ region by using a universal primer to express the domain located on exon 15 in humans. The REJ from synthetic DNA was cloned by using the phosphoramidite synthetic method. Sequencing technology and bioinformatic tools were applied to confirm the validation of the coding region. The results indicated that we successfully cloned and expressed the REJ protein using the DNA synthesis method instead of the conventional methodology.This is an open-access article distributed under the terms of the Creative Commons Attribution-Non Commercial-Share Alike 4.0 License, which allows others to remix, tweak, and build upon the work non commercially, as long as the author is credited and the new creations are licensed under the identical terms.

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Section: Plasmid Dna Purification and Extractionmentioning

confidence: 99%

Cloning and Expression of Receptor of Egg Jelly Protein of Polycystic Kidney Disease 1 Gene in Human Receptor of Egg Jelly Protein

Sonbol¹,

AlRashidi²

2022

Pharmacophore

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Early Cell Cultures from Prostate Cancer Tissue Express Tissue Specific Epithelial and Cancer Markers

Ryabov

Baryshev

Voskresenskiy

et al. 2023

IJMS

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Prostate cancer (PCa) is a widespread oncological disease that proceeds in the indolent form in most patients. However, in some cases, the indolent form can transform into aggressive metastatic incurable cancer. The most important task of PCa diagnostics is to search for early markers that can be used for predicting the transition of indolent cancer into its aggressive form. Currently, there are two effective preclinical models to study PCa pathogenesis: patients derived xenografts (PDXs) and patients derived organoids (PDOs). Both models have limitations that restrict their use in research. In this work, we investigated the ability of the primary 2D prostate cell cultures (PCCs) from PCa patients to express epithelial and cancer markers. Early PCCs were formed by epithelial cells that were progressively replaced with the fibroblast-like cells. Early PCCs contained tissue-specific stem cells that could grow in a 3D culture and form PDOs similar to those produced from the prostate tissue. Early PCCs and PDOs derived from the tissues of PCa patients expressed prostate basal and luminal epithelial markers, as well as cancer markers AMACR, TMPRSS2-ERG, and EZH2, the latter being a promising candidate to mark the transition from the indolent to aggressive PCa. We also identified various TMPRSS2-ERG fusion transcripts in PCCs and PDOs, including new chimeric variants resulting from the intra- and interchromosomal translocations. The results suggest that early PCCs derived from cancerous and normal prostate tissues sustain the phenotype of prostate cells and can be used as a preclinical model to study the pathogenesis of PCa.

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A new device-mediated miniprep method

Cited by 2 publications

References 27 publications

Cloning and Expression of Receptor of Egg Jelly Protein of Polycystic Kidney Disease 1 Gene in Human Receptor of Egg Jelly Protein

Cloning and Expression of Receptor of Egg Jelly Protein of Polycystic Kidney Disease 1 Gene in Human Receptor of Egg Jelly Protein

Early Cell Cultures from Prostate Cancer Tissue Express Tissue Specific Epithelial and Cancer Markers

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