Hi there,

If your intention is to express eukarya proteins inbacteria then you have to clone cDNAs as splicing doesn't occur in bacteria. You can clone both cDNAs into an operon (ie. under the control of one single bacterial promoter). To do so you have to insert a RBS between the stop codon of the first cDNA and the ATG codon of the second cDNA. As alternative you can consider cloning both cDNA in plasmid containing two different promoters (such as pET-DUET for instance). An other approach is to clone cDNAs into 2 different plasmids bearing 2 different markers for selection and then perform double tranformation.

As Dominique responded, cloning should not be a problem. The main question is how you want to express these two genes. If you want to co-express the two genes (to be more precise, the two open reading frames), you can simply clone them into a poly-cistronic vector such as pST39. If you want the two to express independently, you can put them under two separate promoters.

Dear Koti...,

Dual expression vectors for bacteria can be purchased from Novagene (now Merck). You have a choice of high copy (pRSFDuet and pETDuet) or low/medium copy (pACYCDuet and pCDFDuet) vectors and expression strains to regulate the amount of protein being expressed. As mentioned before, you have to have cDNA cloned in because there is no splicing of RNA in bacteria.

Polycistronic vectors can be easily constructed in-house. I would suggest getting a commercial vector since you do not have enough experience in genetic manipulation to carry it out without a snag.

Best regards,

Wes