Published October 29, 2021 | Version v1
Journal article Open

Extraction and reverse transcription of total rna from mouse brain-derived endothelial cells.3 infected by streptococcus Suis 2

Authors/Creators

  • 1. Sumy National Agrarian University

Description

Streptococcus suis 2 is an important emerging zoonotic pathogen. It mainly causes meningitis in pigs. We use SS2 to infect bEnd.3 to get stable cDNA for next research on differences in gene expression and protein expression of cytokines. The paper presents an SS2 study for bEnd.3 infection to obtain stable cDNA for subsequent study of differences in gene expression and cytokine protein expression. Objective: The aim of this study was to extract the total RNA from mouse brain-derived Endothelial cells (bEnd.3) infected by Streptococcus suis serotype 2 (SS2) and transcript to complementary DNA (cDNA). Materials and methods: SS2 strain were obtained from Jilin University, China. BEnd.3 was from Henan institute of Science of Technology, China. Reverse transcription kit was from Takara company, Japan. Trizol was from Bioteke company,China. Nanodrop instrument was from Thermo company, USA. Polymerase chain reaction (PCR) instrument was from Biometra company, Germany. We used SS2 to infect bEnd.3 at a multiplicity of infection (MOI) of 100 for 12h. Cells were harvested and Trizol method was chose to extract the total RNA of bEnd.3 infected by SS2. Nanodrop instrument was used to measure the concentration of RNA and the values of OD260/280 and OD260/230. RNA were transcripted to cDNA with reverse transcription kit by PCR instrument. Results: trizol method used in this study was reliable and high-quality RNA were obtained. Stable cDNA were obtained by reverse transcription kit. Conclusion: in this experiment high-quality RNA was obtained and reverse transcribed to stable cDNA for subsequent detection of related cytokines. This study provides an approximate RNA extraction method and good experimental foundation for downstream research.

Files

Extraction and reverse transcription of total rna from mouse brain-derived endothelial cells.3 infected by streptococcus Suis 2.pdf

Files (640.9 kB)

Additional details

References

  • Liu, F., Li, J., Yan, K., Li, H., Sun, C., Zhang, S. et. al. (2017). Binding of Fibronectin to SsPepO Facilitates the Development of Streptococcus suis Meningitis. The Journal of Infectious Diseases, 217 (6), 973–982. doi: http://doi.org/10.1093/infdis/jix523
  • Dutkiewicz, J., Zając, V., Sroka, J., Wasiński, B., Cisak, E., Sawczyn, A. et. al. (2018). Streptococcus suis: a re-emerging pathogen associated with occupational exposure to pigs or pork products. Part II – Pathogenesis. Annals of Agricultural and Environmental Medicine, 25 (1), 186–203. doi: http://doi.org/10.26444/aaem/85651
  • Coureuil, M., Lécuyer, H., Bourdoulous, S., Nassif, X. (2017). A journey into the brain: insight into how bacterial pathogens cross blood–brain barriers. Nature Reviews Microbiology, 15 (3), 149–159. doi: http://doi.org/10.1038/nrmicro.2016.178
  • Sarkar, A., Fatima, I., Mohammad Sajid Jamal, Q., Sayeed, U., Kalim A. Khan, M., Akhtar, S. et. al. (2017). Nanoparticles as a Carrier System for Drug Delivery Across Blood Brain Barrier. Current Drug Metabolism, 18 (2), 129–137. doi: http://doi.org/10.2174/1389200218666170113125132
  • Sui, Y., Zhang, Z., Wang, H. et. al. (2017). Effects of Suilysin on Apoptosis of Human Cerebral Microvascular Endothelial Cells. Letters in Biotechnology, 3 (28), 290–294.
  • Lin, Z., Zhang, H., Jie, Y. et al. (2020). Modified Trizol Method for Extracting RNA from Mouse Skin. Journal of SUN YAT⁃SEN University, 41 (2), 224–232.
  • George, A. (2018). Simple and efficient method for functional RNA extraction from tropical medicinal plants rich in secondary metabolites. Tropical Plant Research, 5 (1), 8–13. doi: http://doi.org/10.22271/tpr.2018.v5.i1.002
  • Liang, Y., Huang, L., Liu, T. (2021). Corrigendum: Development and Delivery Systems of mRNA Vaccines. Frontiers in Bioengineering and Biotechnology, 9, 718–728. doi: http://doi.org/10.3389/fbioe.2021.766764
  • Fang, T., Xu, Q., Jin, A. et. al. (2021). Comparison of total RNA extraction quality from different tissues of moso bamboo. Journal of Central South University of Forestry & Technology, 9 (41), 157–165.
  • Guo, L., Bai, F., Sha, X. et. al. (2021). Comparison of RNA extraction methods from tomato leaves and fruits. Hubei Agricultural Sciences, 60 (3), 141–144.