I have tried to transfer the plasmids (pGO7 and pMSP353H3) in to L. lactis SMBI 198, and selected in 10 micro/ml con of erythromycine, i got 3-5 colonies and i induced them with 20, 50 and 100 ng/ml of nicin in to GM17 media, after different time interval (2,4...24 hrs) i had pelleted the cells and resuspended into PBS and try to visualised into flurosecent microscope but i could not find any cells containing GFP, i have also used refrence E. coli strain which has some other plasmid having GFP and it give very good fluorocence. I do not know why it happens, can you guide me regarding the same and if you have good protocol for L. lactis transformation could you please send it by mail, i will very thankful to you.
And also i would like to know how we can check the blue white screening in L. lactis, whether it can be done after induction or directly we have to plate them in to X-gal plate,
Please help me
Regards
preeti
0
No replies to this topic
