pBROAD2

pBROAD2 plasmids were designed for the expression of your favorite gene in virtually all tissues of new transgenic mice and rats. pBROAD2 plasmids feature the ubiquitous human ROSA26 promoter, a high CpG content and TATA-less promoter. It directs in vivo expression of the transgene in most tissue types including cells of the developing germ line and hematopoietic lineage. It is also very effective in vitro in a very broad range of mammalian cell lines.

Specifications

- Strong and Ubiquitous Promoter
- Features Multiple Cloning Site or Synthetic LacZ Reporter Gene
- Easy Excision of the E. coli Region
- Efficient in vivo Arrest of the Transgene Transcription

Contents

pBROAD2 plasmids are provided as 20 µg of lyophilized DNA. Plasmids are shipped at room temperature. Store at -20°C.

pBROAD2 plasmids are also available in a kit that contains:
- 20 µg pBROAD2-mcs plasmid
- 20 µg pBROAD2-LacZ containing the lacZ∆CpGnls gene
 

Description

Strong and Ubiquitous human ROSA26 Promoter
This high CpG content promoter directs in vivo expression of the transgene in most tissue types including cells of the developing germ line and hematopoietic lineage. The ROSA26 promoter, initially identified by random retroviral gene trapping in mouse embryonic stem cells [1], directs expression of reporter [2] and recombinase genes [3] in all cells throughout embryonic development and in adult tissues. This TATA-less promoter is very effective in vitro in a very broad range of mammalian cell lines. The strength of the ROSA26 promoter is ascribed to the 10 potential Sp1 sites found within the CpG island extending from the proximal promoter to the the first half of intron 1, the highest number of Sp1 sites ever recorded in any natural promoter.

Multiple Cloning Site or Synthetic Reporter Gene
pBROAD2 is available with a multiple cloning site (MCS) containing several commonly used restriction sites, for convenient cloning of your gene of interest. pBROAD2 MCS 5‘- Age I, Nco I, Bgl II, Hind III, Sal I, Eco RV, Xba I -3‘
pBROAD2 plasmid is also available with the synthetic lacZ∆CpGnls gene, a CpG-free allele of the LacZ gene with a nuclear localization signal.

Easy Excision of the E. coli Region
The E. coli region can be easily removed by using the potent restriction enzyme Pac I. This enzyme recognizes a 8 bp sequence which is located on both sides of the E. coli region.

Efficient in vivo Arrest of the Transgene Transcription
This feature is conferred by the 3‘UTR and polyadenylation sequence of the human β-globin gene [4].

1. Zambrowicz BP. et al. 1997. Disruption of overlappin transcripts in the ROSA beta geo 26 gene trap strain leads to widespread expression of beta-galactosidase in mouse embryos and hematopoietic cells. Proc Natl Acad Sci USA. 94:3789-94
2. Kisseberth WC. et al. 1999. Ubiquitous expression of marker transgenes in mice and rats. Dev Biol.214:128-38.
3. Farley FW. et al. 2000. Widespread recombinase expression using FLPeR (flipper) mice. Genesis. 28:106-10
4. Yu J, Russell JE. 2001. Structural and functional analysis of an mRNP complex that mediates the high stability of human beta-globin mRNA. Mol Cell Biol. 21(17):5879-88