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目的：为了研究人ｅｒａ基因这一人类新基因表达产物的组织分布情况．方法：根据人ｅｒａ序列设计并合成扩增引物，用ＰＣＲ法从ｐＵＣ１９－ｈｅｒａ质粒中扩增人ｅｒａ基因（ｈ－ｅｒａ）全长ｃＤＮＡ和ｈ－ＥＲＡＣ端的结构域基因，并分别克隆到（Ｈｉｓ）６融合表达载体ｐＲＳＥＴ－Ｃ和非融合表达载体ｐＤＨ中，诱导表达（ＨＩＳ）６－ｈ－ＥＲＡ 融合蛋白和 ｈ－ＥＲＡＣ端结构域蛋白．用 ｈ－ＥＲＡＣ端结构域蛋白免疫新西兰白兔，制备兔抗 ｈ－ＥＲＡ抗血清．用 Ｗｅｓｔｅｒｎ－ｂｌｏｔ对兔抗ｈ－ＥＲＡ 抗血清．进行了鉴定，并成功地制备了兔抗ｈ－ＥＲＡ血清．利用在大肠杆菌中高表达、纯化的（Ｈｉｓ）６－ｈ－ＥＲＡ融合蛋白，对已制备的兔抗人 ＥＲＡ抗血清进行进一步鉴定纯化获得抗 ｈ－ＥＲＡ的特异性多抗；再对 ４月～５月人胎儿心、肝、肺、脾、肾等脏器和胃、小肠、睾丸、胸腺、胰脏及胆囊进行免疫组织化学染色．结果：在人类４月～５个月胎儿心、肝、肺、脾、肾脏等脏器和胃、小肠、睾丸中均有免疫反应阳性产物出现．胰脏、心、肺脏等是高表达，脾脏、胃、小肠是中等表达，睾丸、肝脏、肾脏呈低表达，胆囊、胸腺无免疫阳性表达产物出现．结论：人 ＥＲＡ在人类正常胎儿不同组织中?
OBJECTIVE: To study the tissue distribution of the human era gene, a novel human gene product. Methods: Amplification primers were designed and synthesized according to the human era sequence. The full-length cDNA of h-era and the domain of h-ERAC were amplified by PCR from pUC19-hera plasmid and cloned into (His ) 6 fusion expression vector pRSET-C and the non-fusion expression vector pDH induced expression (HIS) 6-h-ERA fusion protein and h-ERAC terminal domain protein. Rabbit anti-h-ERA antiserum was prepared by immunizing New Zealand white rabbits with h-ERAC terminal domain protein. Rabbit anti-h-ERA antiserum was detected by Western-blot. Were identified and successfully prepared rabbit anti-h-ERA serum. The purified anti-human ERA antiserum was further identified and purified by using (His) 6-h-ERA fusion protein highly expressed in Escherichia coli to obtain specific anti-h-ERA polyclonal antibody. May human fetal heart, liver, lung, spleen, kidney and other organs and the stomach, small intestine, testis, thymus, pancreas and gallbladder for immunohistochemical staining. Results: In April ~ 5 months in human fetal heart, liver, lung, spleen, kidney and other organs and stomach, small intestine, testis were immunoreactive products appear. Pancreas, heart and lungs were highly expressed. Spleen, stomach and small intestine were moderately expressed, while testis, liver and kidney were low, while gallbladder and thymus were not immunopositive. Conclusion: Human ERA in different human normal fetal tissues?