Fig. 2.

Nucleotide and amino acid sequence of rabbit CCT-alpha cDNA. Cloning of the CCT-alpha was accomplished as follows: the primer pairs for CCT-alpha listed in Table 1 were used to generate an RT-PCR amplimer product (as shown in Fig. 1). The nucleotide sequence of this product was then determined. This obtained sequence confirmed homology to mouse and human CCT-alpha sequences on the Basic Local Alignment Search Tool search and was then used to design both forward and reverse primers for 3′ and 5′ RACE, respectively. The primer used for 3′ RACE was 5′-GGACCAAATTAGGCAGAGAGAATC-3′. The primer used for 5′ RACE was 5′-GCCGGTAGCCACTGATGACTGA-3′. 5′ and 3′ RACE reactions were carried out as described in the GeneRacer kit protocol (Invitrogen Corporation, cat. no. L1500-1). RACE amplimers of the expected molecular weight were subcloned by Topo TA ligation into the vector provided; the sequence of these derived amplimers was then determined, and the composite whole CCT-alpha cDNA was assembled therefrom. The full-length nucleotide sequence for the rabbit CCT-alpha cDNA is shown, with the corresponding amino acids in the coding region. The start codon (ATG) and stop codon (TGA) are enlarged and bolded. A consensus polyadenylation signal sequence at 1,854 bp (ATTAAA), appropriately positioned 23 bp upstream of the polyA tail, is similarly enlarged and bolded