Vectors for inducible expression and inducible RNAi

Basic pUC vectors

Expression is inhibited by a spacer between promoter and gene of interest. The spacer contains a lox site at the beginning and at the end. The spacer can be removed by Cre recombinase. Expression of Cre recombinase is induced by the Heat Shock Promomoter from Glycine max. After induction you get full expression of your gene of interest.

We have developed two different vectors as examples. One for protein expression and one for RNAi. The structure of both vectors is very similar.

  1. Promoter (P35s) flanked by KpnI and SalI
  2. Marker (NptII with intron) flanked by Lox sites and SpeI and MCS
  3. Spacer (gus-fragm.) flanked by two MCSs
  4. Terminator(Tnos) or second promoter (P35s) flanked by NotI and a MCS
  5. SfiI-A and SfiI-B sites for cloning into the binary

 

pD1-35S-RNAi-NptLox-Nhe genebank sequence image
pD1-35S-NptLox-MCS genebank sequence image

 

Binary vector for cloning of basic pUC vectors

The binary vector contains the Cre recombinase that is under control of the soybean Heat Shock Promoter. At the moment there is only one binary available. If you need another binary we can construct them on request.

p7i-HSP-Cre genebank sequence image

 

pUC vectors with Cre already integrated - you can use any binary with SfiI restriction sites

pD1-35S-Hsp-Cre-RNAi genebank sequence image
pD1-35S-HCE genebank sequence image