Three xenotropic retroviruses have been biologically cloned from cells cultured from the kidney of a 3-month-old NZB female mouse. They were obtained by first cocultivating the kidney cells for several weeks with mink, dog, and human cells and then cloning them by endpoint dilution. The cloned viruses differ in their infectivity and replicative ability in a variety of heterologous cell lines. The mink cell line-derived virus (X-NZB/K-Mlc) reaches titers in culture of over 10(8) infectious viruses/ml, and is produced in high titer within 24 hr after infection of mink lung cells. The human and dog cell-derived NZB viruses (X-NZB/K-Huc and X-NZB/K-Dgc) grow to lower titers and are similar in many respects. They differ in their relative ability to replicate in dog and human cells and to transform mink S+L- cells. Peptide mapping studies indicate that the X-NZB/K-Mlc virus has a unique p15(E) protein which distinguishes it from the other two cloned NZB viruses. These results lend further support to the observation that several types of xenotropic virus are present in a mouse strain and that more than one virus can be expressed by one organ of a particular mouse.