I have cloned a sequence of bacterial gene in pet25b+ vector in DH5alpha strain. For expression, E.coli BL21 cell line was used. However, the gene sequence cloned was 100% correct confirmed via Sanger sequencing.

But unfortunately, protein sequence calculated from NMR experiment of doubly labeled protein is showing additional amino acid residues in comparison to NCBI provided sequence. Could anyone help me in this issue?

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