Genetic instability of Japanese encephalitis virus cDNA clones propagated in Escherichia coli

摘要: The genetic instability of Flavivirus cDNA clones in transformed bacteria is a common phenomenon. Herein, fragment the nucleotide (nt) 1–2913 genome flavivirus, Japanese encephalitis virus (JEV), was used to investigate factors that caused clones. Several fragments with different 5′- or 3′-termini 2913-nt were obtained by PCR amplification restriction enzyme digestion and cloned into pCR-Blunt II-TOPO vector. All stably propagated at 25 °C. However, 5′-untranslated region half 3′-E gene could cause 37 °C. 5′-terminus sequences subjected testing prokaryotic promoter activity luciferase assay Western blot. 54–120 nt JEV exhibited high 37 °C, declined markedly These findings revealed together expression structural genes determined clone. Growth room temperature may reduce 5′-sequences represent an effective way improve stability flavivirus host bacteria.