Atp11c基因诱捕小鼠遗传背景分析

目的利用PCR(polymerase chain reaction,PCR)和荧光竞争性PCR技术对构建的Atp11c基因诱捕小鼠的遗传背景进行分析。方法通过PCR技术鉴定基因诱捕载体的插入位点和宿主染色体的缺失状态;利用荧光竞争性PCR技术检测宿主染色体内的诱捕载体拷贝数。结果 54对引物的PCR结果确定了诱捕载体在Atp11c第一个内含子中的插入位点,测序结果显示伴随着基因诱捕载体两端大于200 bp的碱基缺失,宿主染色体片段出现了418 bp的碱基删除;荧光竞争性PCR证实诱捕载体为单拷贝整合。结论成功解析了Atp11c基因诱捕小鼠的遗传背景,建立了快速、准确检测诱捕小鼠遗传背景的方案。

Analysis of the genetic background of Atpllc gene trap mice

Objective To analyze the genetic background of Atpl lc gene trap mice. Methods The entire re- gion PCR was applied to find the gene trap vector insertion site and the lack state of the host chromosome. Fluorescent com- petitive PCR method was used to detect the copy numbers of the integrated vector（s）. Results The PCR results by 54 pairs of primers showed that the trapping vector insertion site was in the first intron of Atpllc gene, and the DNA sequen- cing results displayed 418 bp deletion on the host chromosome and more than 200 bp missing on both ends of the vector. Fluorescent competitive PCR confirmed a singte copy trapping vector inserted. Conclusion The genetic background of Atpl l c gene trap has been resolved successfully and a program has been established to detect the genetic background of gene trap mice accurately and rapidly.