Gene Cloning and Prokaryotic Expression Vector Construction of Alpha-Lactalbumin One of the Major Allergens in Milk

The RT-PCR was applied to clone the gene of alpha-lactalbumin and the specific primers were designed to amplify the ORF of the alpha-lactalbumin gene sequence.Insert the ORF fragment into the Expression Vector PET-28A and construct the relevant prokaryotic Expression Vector.Succeed to clone the gene of alpha-lactalbumin the major allergen in milk and construct the revelant prokaryotic Expression Vector ultimately.The cloned cDNA ORF sequence contained 429 bp and encoded 142 amino acids.Sequence analysis shows the estimated molecular mass 16 265 and pI 6.10 and this clone has the high homology with the alpha-lactalbumin sequences of bovine submitted on the Genebank.Therefore,this gene sequence is regarded as the bovine alpha-lactalbumin gene(GenBank database entry No.FJ232912).This study will be the basic for the expression of the allergen gene and the further research.