Figure 1

Amplification of 185/333 sequences. A. gDNA from animal 2 was amplified by PCR using primers flanking the 185/333 genes (185-5'UTR and 185-3'UTR; see Table 1) and showed bands of five major sizes from 1.2 to 2 kb and a minor band around 4 kb. Triangles indicate the positions of each of the bands. Similar amplification of gDNA from six additional animals had identical patterns of bands (data not shown). B. Amplification of intergenic regions by PCR using primers that annealed within genes but were oriented away from each other (185-LR1 and 185-F5 primers; Table 1) revealed a major band in all animals of about 3 kb (lanes 1–4, animals 10–13). C. gDNA from animal 2 was amplified by PCR using primers that annealed in the 5' UTR (185-5' UTR; Table 1) and the type I repeats found in elements Ex4, Ex5, and Ex6 (185-R5; Table 1) and revealed the presence of genes that lacked introns. Animal 2 gDNA (lane 1) shows a band of 450 bp (indicated with a carrot). A cDNA (Sp0313, DQ183171; lane 2) also amplifies a band of about 450 bp. A cloned gene (2-02, Additional file 1; GenBank accession number EF607673; lane 3) with a typical intron amplifies a band within the range of bands in A. D. PCR amplification of 185/333 from eight different BAC clones using primers (185-5'UTR and 185-3'UTR; Table 1). Each lane contains template DNA from a different BAC (1, 126J14; 2, 108H07; 3, 053M03; 4, 121A07; 5, 004D19; 6, 182N21; 7, 148J22; 8, 019L13).