中文摘要：

目的：采用载体构建及荧光定量PCR的方法比较BCL2等位基因不同部位PCR引物在定量分析中的扩增效率,判断其用于不同等位基因转录活性分析的可靠性.方法：用PCR扩增包含定量PCR目的片段的序列,然后将扩增产物分别插入pGEM-T Easy载体中,构建TA3重组载体,再以TA3载体为模板,采用荧光定量PCR方法比较引物对之间扩增效率.结果：定量PCR结果显示引物对之间扩增效率无明显差异.结论：所检测的PCR引物对之间的扩增效率一致,可用于比较分析mbr^＋/mbr^-Nalm-6杂合子细胞系中两个等位基因表达活性.

英文摘要：

Objective： To investigate the amplification efficiency of primers on different sites of BCL2 alleles in real-time PCR reactions and the reliability of these primers in analysis of transcription activity of different alleles with vector construction and real-time PCR. Methods： Fragments containing all amplicons from real-time PCR reactions wereamplified by PCR and then inserted into the pGEM-T Easy vector to construct TA3 vector. The real-time PCR analysis was performed with TA3 plasmid as templates for testing the amplification efficiency of primers. Results： Real-time PCR showed no significant difference in the amplification efficiency for the of primers. Conclusion： Amplification efficiency of primers in real-time PCR reactions is identical. These primers can be used for quantity analysis of transcription activity of two BCL2 alleles in mbr^＋/mbr^-Nalm-6 heterozygous cell line.