ABSTRACT

The Tg(Adipoq-cre)1Evdr mouse has become an important tool in adipose tissue biology. However, the exact genomic transgene integration site has not been established. Using Targeted Locus Amplification (TLA) we found the transgene had integrated on mouse chromosome 9 between exons 6 and 7 of Tbx18. We detected transgene-transgene fusion; therefore, we used droplet digital polymerase chain reaction to identify Cre copy number. In two separate experiments, we digested with BAMHI and with HindIII to separate potentially conjoined Cre sequences. We found one copy of intact Cre present in each experiment, indicating transgene-transgene fusion in other parts of the BAC that would not contribute to tissue-specific Cre expression. Cre copy number for Tg(Adipoq-cre)1Evdr mice can be potentially used to identify homozygous mice.

 抽象的

Tg(Adipoq-cre)1Evdr 小鼠已成为脂肪组织生物学的重要工具。然而，确切的基因组转基因整合位点尚未确定。使用靶向基因座扩增 (TLA)，我们发现转基因已整合到小鼠 9 号染色体上Tbx18外显子 6 和 7 之间。我们检测到转基因-转基因融合；因此，我们使用液滴数字聚合酶链式反应来鉴定Cre拷贝数。在两个单独的实验中，我们用 BAMHI 和 HindIII 消化以分离可能相连的Cre序列。我们在每个实验中都发现了一份完整的Cre副本，表明 BAC 其他部分中的转基因-转基因融合不会有助于组织特异性Cre表达。 Tg(Adipoq-cre)1Evdr 小鼠的Cre拷贝数可潜在用于鉴定纯合小鼠。