Does anybody knows any method to enhance the recombination of genes to transform industrial S. cerevisiae?

Enrique Javier Carvajal Barriga @Enrique_Carvajal_Barriga

11 November 2011 1 4K Report

I've got a 9.4 kb construct to be recombined in ribosomal genes. I have tried a number of times to recombine this construct, but I haven't been succesful.

Thanks

Enrique Javier Carvajal Barriga

Thanks for your response. I tried to transform industrial S. cerevisiae strains by competent cells method and spheroplasts. The construct is designed with a geneticin resistance gene and other genes to be expressed. The construct is flanked with 40bp sequences to recombine in ribosomal genes. I tried several times to recombine this construct, but I didn't succeed. What I suspect is that the construct is too large and is probably making hairpins, so I need to find a way to enhance or facilitate recombination of this exogenous DNA in the yeast chromosome.

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