I wish to clone a gene (mammalian gene) in a plasmid and express it in bacteria. I need the features for the cloned protein: 1. His tagged (for purification purpose) 2. GFP (to visualize) 3. Secreted (harvest from bacterial culture supernatant). I would like to know which vector allows me to have these features. Please suggest me.
Thank you
I solved my question and thought to share with others. A new system from Promega "Flexi vector" is very nice. A high affinity HQ-tag is a nice feature in it. Comes in amp+ and kan+ options for selections. I did Spliced Overlapping PCR (SOP) with my gene of interest and GFP and then cloned into Flexi vector. Transformed into JM109 as recommended by Promega. It worked straight away. I hope this will help others.
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