I want to expression a gene from actinomycete. My gene's size is around 800 bp. The Restriction enzymes are BamHI and HindIII. I isolated and purified my gene and ligated into pET28 ( pEt28 was digested by BamHI and HindIII too), then i transformed in XL1blue, kept plate overnight. I got around 30 colonies, i selected 10 and isolated, digested but i could not get any band of gene. i have repeated 2 times, but all are same.
Could someone know what is my problem? please help me, im waiting for your answers. Thank you so much. My english is not good, im so awkward, plz don't mind ^^