通过实时定量PCR对V(D)J重组进行定量分析。

Quantification of V(D)J recombination by real-time quantitative PCR.

作者信息

Braikia Fatima-Zohra, Chemin Guillaume, Moutahir Mohammed, Khamlichi Ahmed Amine

机构信息

CNRS UMR 5089; IPBS (Institut de Pharmacologie et de Biologie Structurale); FRBT, 205 route de Narbonne, BP 64182, F-31077 Toulouse, France; Université de Toulouse; UPS; IPBS F-31077 Toulouse, France.

出版信息

Immunol Lett. 2014 Nov;162(1 Pt A):119-23. doi: 10.1016/j.imlet.2014.08.002. Epub 2014 Aug 12.

DOI:10.1016/j.imlet.2014.08.002

PMID:25128366

Abstract

B and T lymphocytes have the unique capacity to somatically rearrange their antigen receptor loci through V(D)J recombination. D-JH and VH-DJH recombination events are usually visualized by semi-quantitative PCR followed by detection of end products, which is time consuming and requires the use of hazardous elements. Additionally, it necessitates relatively large amounts of genomic DNA which could be limiting when the cell populations of interest are rare. Here, we describe a real-time quantitative PCR assay for a fast quantification of V(D)J recombination events at the IgH locus.

摘要

B淋巴细胞和T淋巴细胞具有通过V(D)J重组对其抗原受体基因座进行体细胞重排的独特能力。D-JH和VH-DJH重组事件通常通过半定量PCR，随后检测终产物来可视化，这既耗时又需要使用危险元素。此外，它需要相对大量的基因组DNA，当感兴趣的细胞群体稀少时，这可能会受到限制。在此，我们描述了一种实时定量PCR检测方法，用于快速定量IgH基因座处的V(D)J重组事件。