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. Author manuscript; available in PMC: 2017 Oct 1.
Published in final edited form as: J Cardiovasc Electrophysiol. 2016 Aug 30;27(10):1220–1229. doi: 10.1111/jce.13049

Figure 1.

Figure 1

pMHC-TGF-β1cys33ser knock-in vector construction. The pcDNA3.1D V5 vector was linearized by digestion with Bgl II, treated with Klenow to blunt the ends, followed by cutting with Xho I; these treatments resulted in the linearized pcDNA3.1D V5 vector bearing a blunt end at the Bgl II cut site and a sticky end generated by Xho I digestion. The MHC-TGF-β1cys33ser cassette was liberated from vector pUC-BM20-MHC-TGF-β1 by double digestions with Nru I and Sal I. As Nru digestion generates blunt ends and Sal I digestion generates a compatible sticky end to the sticky end generated by Xho I, the MHC-TGF-β1cys33ser cassette was directly subcloned into the pcDNA3.1DV5 vector prepared above to obtain the pMHC-TGF-β1cys33ser knock-in vector.