An apparatus and method is proved that transmits and receives Reference Signals (RSs) in a wireless communication system using Multiple-Input and Multiple-Output (MIMO). A system adapted to the method is provided. The method includes: determining the number of RS transmission to two or more antennas, respectively, as the number of antennas included in respective ports, wherein each port include one or more antennas; creating transmit patterns so that they cyclically differ from each other every the number of RS transmission to transmit the created transmit patterns the number of RS transmission times; and mapping the RSs to one or more antennas according to the created, respective patterns, and transmitting the RSs in order.