This plasmid contains an insulin-cre-hGH fusion gene that was made as follows. (1) The 1.2 kb NLSKOZCre gene from pmL78 was subcloned into pBSIIKS at Sal I and EcoRI. (2) The 0.7 kb rat insulin promoter 2 gene (RIP) was subcloned from pSPRIP into the Kpn I and Xho I sites. (3) The 2.1 kb growth hormone gene from pOGH was cut at BamHI and EcoRI, blunted, and Spe I sites introduced on either end; this GH gene fragment was then subcloned into the Spe I site of the clone from step 2. (4) A small Sal I/Sal I polylinker fragment between RIP and Cre was deleted. (5) The final construct (pBS/RIP-Cre-GH) was cut at Kpn I and Not I and used for microinjection. This plasmid was used to generate RIP(Ins2)-Cre transgenic mice.


Category
Plasmid
Record History
Added on December 23, 2008 at 9:13 AM by Gangula, Rama
Modified on October 2, 2015 at 4:32 PM by Magnuson, Mark
Shared with (contributions)
Public: Shared Collection

Vector Annotations

Vector Map AeXNzHDz.png
Genbank File pRIP-Cre-GH.gb
Backbone Vector pBluescript II KS
Construct Size 6,936 bp (approximate)
Bacterial Stock No
Storage Temperature -20°C
Stock Concentration 0.1 µg/µL
Addgene Not Provided

Source

Laboratory Mark Magnuson
Made byYuejin Chen
Stock Date December 23, 2008

Inventory Location

Private

Publications / Citations

  1. Dual roles for glucokinase in glucose homeostasis as determined by liver and pancreatic beta cell-specific gene knock-outs using Cre recombinase. Postic C, Shiota M, Niswender KD, Jetton TL, Chen Y, Moates JM, Shelton KD, Lindner J, Cherrington AD, Magnuson MA (1999) J Biol Chem 274(1): 305-15
    › Primary publication · 9867845 (PubMed) · Added on 1/31/2014

MeSH Terms

Albumins Alleles Animals Base Sequence DNA Primers Gene Deletion Glucokinase Glucose Homeostasis Insulin Integrases Islets of Langerhans Liver Mice Mice, Knockout Transgenes Viral Proteins