Abstract: Nicotinic acetylcholine receptors (nAChRs), an important kind of ligand-gated ion channels with many subtypes, are similar in structures but different in physiological and pharmacological functions. Natural α6β2^* nAChRs are unlikely to be expressed in vitro and their expression currents are quite small. In order to establish an effective experimental model of α6/α3β2β3 nAChRs in vitro, the expression system of Xenopus laevis oocytes was used to express the α6/α3β2β3 nAChRs subtype with the method of micro injection of RNA and detection with two-electrode voltage clamp. The expression conditions were optimized to obtain a better and more efficient expression system for α6/α3β2β3 nAChRs. The current generated by the expressed receptor was significantly increased. This may provide an experimental model and basis for the screening of new drugs targeting this subtype.