In the pond ciliate Tetrahymena thermophila, expression of genes encoding variant forms of the cell surface immobilization antigen (i-ag) is regulated by environmental conditions. Multiple isoforms of the L i-ags are found on the surface of cells grown at <20C as well as on the surface of rseC mutants which express SerL genes constitutively. Five cDNAs encoding variant L i-ags of rseC were sequenced and their expression studied. Two additional SerL genes from natural isolates were sequenced. Members of the SerL family encode polypeptides with 148, 316, or 371 amino acids, and the i-ags have two, five, or six imperfect repeats, respectively, flanked by putative ER translocation and GPI addition signals. Each repeat contains six periodic cysteines, in contrast to eight or ten in other i-ags of T. thermophila. At least three of the five genes constitutively expressed in rseC mutants are differentially expressed in cells expressing other i-ags. Northern analysis and RT-PCR indicate that expression of some members of the SerL family is regulated by both transcription and mRNA stability while another member is regulated primarily by mRNA stability.