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用新鲜的人绒毛膜组织,抽提组织总RNA并通过逆转录反应合成cDNA第一链。利用人工合成的一对寡核苷酸引物,采用PCR技术特异性地扩增hCG-β-cDNA。琼脂糖凝胶电泳结果显示扩增片段大小为500bp,同预计的片段长度相符。将此片段利用T-A载体克隆至PCRⅡ质粒上并进行全序列分析,结果显示克隆片段包含hCG-βcDNA5’端和3’端非编码区及完整的编码区。
With fresh human chorionic tissue, total tissue RNA was extracted and cDNA first strand was synthesized by reverse transcription reaction. HCG-β-cDNA was specifically amplified by PCR using a pair of synthetic oligonucleotide primers. The results of agarose gel electrophoresis showed that the amplified fragment size was 500bp, which was consistent with the predicted fragment length. The fragment was cloned into the PCR II plasmid using the T-A vector and subjected to full sequence analysis. The results showed that the cloned fragment contains the 5 ’end and the 3’ end non-coding region and the complete coding region of the hCG-β cDNA.