Amplification of full-length transcripts for WC1s. (A) Primers designed for complete coding sequences of all thirteen WC1 genes. Schematic representations of the molecular forms of archetypal WC1 genes and WC1-11 with primer placement indicated. The WC1 common forward primer (WC1atg-for) for complete coding sequences was designed based on the conserved region in the signal sequences, while the reverse primers (WC1group1,2-rev and WC1group3-rev) were based on the end of the 3′ coding sequences. Abbreviations are as follows: ID, inter-domain sequence; TM, transmembrane region; ICD, intracytoplasmic domain. (B) cDNA evidence for WC1 genes. Primer pairs WC1atg-for/WC1group1,2-rev (designed for WC1group1,2) and primer set WC1atg-for/WC1group3-rev (designed for WC1group3) were used to amplify all the complete coding sequences of WC1 transcripts as described in the previous study . (C) Confirmation of complete coding sequences for WC1-nd1 and WC1-nd2. Four different templates used in PCR for all thirteen WC1 domain 1 specific primer pairs are indicated in the left part of each gel. (D) Agarose gel electrophoresis evidence for complete coding sequences of WC1-nd1 and WC1-nd2. Complete coding sequences of WC1-nd1 and WC1-nd2 amplified by primer pairs of specific forward primers and common reverse primers (WC1group1,2-rev). The cDNA isolated from sorted WC1.1+ γδ T cells was used as a template. Gel electrophoresis of the PCR products was performed on 1% agarose gel. For each primer set, the identity of the amplified products was confirmed by DNA sequencing analysis.
Chen et al. BMC Genetics 2012 13:86 doi:10.1186/1471-2156-13-86