We investigated the effect of long term passage of parasites in naive animals on the circumsporozoite protein (CSP) gene of Plasmodium yoelii. The CSP gene sequence was determined from a non-lethal cloned line of P. yoelii and compared to the CSP gene sequence from a lethal strain of P. yoelii. The two parasite lines were originally derived from the same isolate, but were separated 17 years ago followed by continued passage. The sequence of the CSP gene and its surroundings from the non-lethal line remains identical to that from the lethal isolate except for a deletion within the repeated central domain. This result contrasts with the results obtained by sequencing a number of clones from different geographical field isolates of Plasmodium falciparum where there appears to be rapid accumulation of sense mutations within putative functional domains. These observations are consistent with the suggestion that strong biological pressure in a field environment results in selection of parasite types on the basis of different CSP gene sequences.