Li-Hua Li

When expression of more than one gene is required in cells, bicistronic or multicistronic expression vectors have been used. Among various strategies employed to construct bicistronic or multicistronic vectors, an internal ribosomal entry site (IRES) has been widely used. Due to the large size and difference in expression levels between genes before and after IRES, however, a new strategy was required to replace IRES. A self-cleaving 2A peptide could be a good candidate to replace IRES because of its small size and high cleavage efficiency between genes upstream and downstream of the 2A peptide. Despite the advantages of the 2A peptides, its use is not widespread because (i) there are no publicly available cloning vectors harboring a 2A peptide gene and (ii) comprehensive comparison of cleavage efficiency among various 2A peptides reported to date has not been performed in different contexts. Here, we generated four expression plasmids each harboring different 2A peptides derived from the foot-and-mouth disease virus, equine rhinitis A virus, Thosea asigna virus and porcine teschovirus-1, respectively, and evaluated their cleavage efficiency in three commonly used human cell lines, zebrafish embryos and adult mice. Western blotting and confocal microscopic analyses revealed that among the four 2As, the one derived from porcine teschovirus-1 (P2A) has the highest cleavage efficiency in all the contexts examined. We anticipate that the 2A-harboring cloning vectors we generated and the highest efficiency of the P2A peptide we demonstrated would help biomedical researchers easily adopt the 2A technology when bicistronic or multicistronic expression is required.

<i>Aim</i>: Hepatitis B virus is a primary etiological factor for various liver diseases, including cirrhosis and hepatocellular carcinoma.Hepatitis B virus is an incomplete double-stranded DNA virus, which infects hepatic cells, enters the nucleus to form a complete double-stranded DNA and products a series of functional proteins, then replicates and assembles to form a complete virus, which is released outside the cell. So far, the pathogenesis of hepatitis B virus is not clear. JNK signaling pathway is an important branch of MAPK pathway, which plays an important role in various physiological and pathological processes such as cell cycle, reproduction, apoptosis and cell stress. Study show JNK activation involves liver damage. Especially, hepatitis B virus infection promote the phosphorylation of JNK. Chlorogenic acid, as a polyphenolic compound, exhibits notable antioxidant and antiviral properties. Study revealed chlorogenic acid had ability of inhibiting hepatitis B virus replication, but the mechanisms was unknown. Here we demonstrated the antiviral mechanisms of chlorogenic acd on HBV replication. <i>Methods</i>: To investigate the effect of chlorogenic acid on HBV replication, southern blot and western blot were performed using HepAD38 cells. <i>Results</i>: Chlorogenic acid suppressed HBV replication. In this process, JNK expression was inhibited. <i>Conclusion</i>: chlorogenic acid suppressed HBV replication via inhibiting JNK expression.