Raaphorst F M, Tami J, Sanz I E
University of Texas Health Science Center at San Antonio 78284-7874, USA.
Biotechniques. 1996 Jan;20(1):78-82, 84, 86-7. doi: 10.2144/96201st02.
Methods have been developed to rapidly visualize the size distribution of third complementarity-determining regions (CDR3) in immunoglobulin (Ig) and T-cell receptor (TCR) molecules. DNA fragments spanning the Ig or TCR CDR3 are generated by PCR using primers at fixed positions in the variable and constant segments. These fragments differ in length due to size variation of the CDR3s. Visualization of the amplification products in polyacrylamide gels as a "CDR3 fingerprint profile" is a rough measure for the complexity of the Ig and TCR antigen-binding specificities. We report an adaptation of this method for the analysis of human Ig heavy-chain genes that incorporates silver staining, which allows for the fine analysis of specific regions of the profiles. This is especially useful for the study of low-abundant transcripts.
已经开发出一些方法来快速可视化免疫球蛋白（Ig）和T细胞受体（TCR）分子中第三互补决定区（CDR3）的大小分布。使用可变区和恒定区固定位置的引物，通过聚合酶链反应（PCR）生成跨越Ig或TCR CDR3的DNA片段。由于CDR3大小的变化，这些片段长度不同。在聚丙烯酰胺凝胶中将扩增产物可视化为“CDR3指纹图谱”，是衡量Ig和TCR抗原结合特异性复杂性的一种粗略方法。我们报告了一种适用于分析人类Ig重链基因的该方法的改进版本，该版本采用银染法，能够对图谱的特定区域进行精细分析。这对于研究低丰度转录本特别有用。
