Determination of the sequence coding for the β subunit of the human high‐affinity IgE receptor

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Abstract
The cDNA encoding the β subunit of the human high‐affinity IgE receptor was cloned by a combination or various polymerase chain reactions (PCR). A major portion of the β cDNA was amplified using primers homologous within the sequences of rat and mouse. The 3′ unknown sequence was preferentially amplified using the RNA template‐specific PCR and the improved two‐step PCR. The 5′ unknown sequence was specifically amplified by our newly developed PCR walking. Random heptanucleotides tagged with a unique sequence at the 5′ end were used as the walking primer. Finally, the entire coding region was amplified and sequenced. The two extracellular loops of the human β subunit were the least homologous to those of rat and mouse.