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本研究采用逆转录－ＰＣＲ技术克隆自身核抗原Ｕ１ＳｎＲＮＰ７０ｋＤ多肽分子中Ｕ１ＲＮＡ结合功能区（Ｕ１ＲＮＡｂｉｎｄｉｎｇｄｏｍａｉｎ，Ｕ１ＢＤ）的ｃＤＮＡ，经ＤＮＡ测序证实以后定向插入原核表达载体ＰＧＥＸ－２Ｔ，进而导入大肠杆菌中表达重组蛋白。免疫印迹法研究表明：９６％（４８／５０）的抗Ｕ１ＳｎＲＮＰ７０ｋＤ抗体阳性血清能够识别该重组蛋白，证实Ｕ１ＢＤ重组蛋白具有Ｕ１ＳｎＲＮＰ７０ｋＤ抗原性，而且Ｕ１ＢＤ是Ｕ１ＳｎＲＮＰ７０ｋＤ多肽上主要抗原表位区域，能够被大多数抗Ｕ１ＳｎＲＮＰ７０ｋＤ抗体阳性血清识别。这为今后对Ｕ１ＢＤ抗原表位精确定位，分析特定抗原表位与疾病的相关性以及制备重组抗原用于临床检测等研究奠定基础。
In this study, U1 RNA binding domain (U1BD) cDNA of U1SnRNP70kD polypeptide was cloned by RT-PCR and confirmed by DNA sequencing and then inserted into prokaryotic expression vector PGEX-2T, which was then introduced into Escherichia coli to express the recombinant protein . Immunoblotting studies showed that 96% (48/50) of anti-U1SnRNP70kD antibody-positive sera can recognize the recombinant protein, confirming U1BD recombinant protein has U1SnRNP70kD antigenicity, and U1BD is the major antigenic epitope region on U1SnRNP70kD polypeptide, Anti-U1SnRNP70kD antibody-positive serum recognition. This laid the foundation for future research on the precise location of U1BD epitopes, the analysis of the correlation between specific epitopes and diseases, and the preparation of recombinant antigens for clinical testing.