کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
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2821610 | 1160970 | 2009 | 9 صفحه PDF | دانلود رایگان |
![عکس صفحه اول مقاله: Next generation tools for high-throughput promoter and expression analysis employing single-copy knock-ins at the Hprt1 locus Next generation tools for high-throughput promoter and expression analysis employing single-copy knock-ins at the Hprt1 locus](/preview/png/2821610.png)
We have engineered a set of useful tools that facilitate targeted single copy knock-in (KI) at the hypoxanthine guanine phosphoribosyl transferase 1 (Hprt1) locus. We employed fine scale mapping to delineate the precise breakpoint location at the Hprt1b-m3 locus allowing allele specific PCR assays to be established. Our suite of tools contains four targeting expression vectors and a complementing series of embryonic stem cell lines. Two of these vectors encode enhanced green fluorescent protein (EGFP) driven by the human cytomegalovirus immediate-early enhancer/modified chicken beta-actin (CAG) promoter, whereas the other two permit flexible combinations of a chosen promoter combined with a reporter and/or gene of choice. We have validated our tools as part of the Pleiades Promoter Project (http://www.pleiades.org), with the generation of brain-specific EGFP positive germline mouse strains.
Journal: Genomics - Volume 93, Issue 3, March 2009, Pages 196–204