Gene Engineering And Functional Identification Of Hepatitis B Virus Core Protein

Objective To search a gene engineering site and form of HBV coreprotein for forming DNA loading and cell-penetrating peptides or ligandcarried viral like particles (VLPs), which would be a foundation work todevelop HBV infection model.Methods1HBc, HBV1.1c-and all gene engineered HBc were constructed byRestriction Digestion and Ligation-independent Cloning (RLIC).2To determine the function of gene engineered HBc, therecombined HBc constructs were cotransfected with HBV1.1c-intoHEK293cells, and HBV DNA replicative intermediates in core particleswere assayed by Southern blot.Results1In the HEK293cells, WtHBc and HBV1.1c-have functionaltrans-complementation to support HBV replication.2The replacement or deletion of HBc apical loop region of aa79-80failed to support HBV replication.3HBc aa86-93replacement or deletion have functional defect.4The oligopeptide insertion at N-terminal of HBc have the functionto support HBV replication at a low level.5HBc N-terminal can be the functional site for extrinsic proteinfusion, which can support the insertion of EGFP(238aa), RFP(225aa), VSVG(511aa).Conclusion We developed a effective trans-complementation systemof HBc and HBV1.1c-to support HBV replication. This system can be usedto identify the function of gene engineered HBc. Using this system, wefound that HBc functional deficit when the replacement or deletionhappened at aa79-80or aa86-93, and the N-terminal can be the effectivesite for engineering.