Abstract:

In order to establish a bovine lactoferricin biosynthesis method, the bovine lactoferricin-encoding gene was determined on the basis of amino acid sequence. A recombinant fusion gene was obtained by means of overlapping PCR following the purification of the separate PCR products of the bovine lactoferricin-encoding sequence and the split intein Asp dnaEIn gene from the DNA polymerase gene dnaENI of Anabaena sp. PCC 7120, and the obtained fusion gene was subsequently cloned into the expression plasmid pET-28a to construct a recombinant fusion expression plasmid. The results showed that recombinant fusion proteins were over-expressed after the expression host Escherichia coli BL21 (DE3) containing the recombinant bovine lactoferricin fusion expression plasmid was induced and that reaction products of over-expressed recombinant bovine lactoferricin lysates and the purified Anabaena sp. PCC 7120 DnaECI, which contained split intein Asp DnaEIc, could inhibit Staphylococcus aureus.

Key words: bovine lactoferricin, overlapping PCR, fusion gene, fusion protein