Molecular cloning and expression of cDNA for human antileukoprotease from cervix uterus

We have isolated cDNA clones for the human antileukoprotease HUSI-I, an Elastase Inhibitor, from a library, containing cDNA inserts made from human cervix uterus. A library of 10 000 recombinants was screened using a mixture of 16 different oligodeoxyribonucleotides which correspond to Amino acids 79-84 and one 20mer oligodeoxyribonucleotide corresponding to Amino acids 19-26. Two overlapping cDNA clones, containing the entire coding sequence and part of the 5'- and 3'-untranslated region, were isolated. DNA sequence data showed that our clone corresponds with the available protein sequence data. For expression, the cDNA fragment was inserted in a derivative of plasmid pPLc236 and expressed under the control of lambda PL promoter. Expression of antileukoprotease was proven by Western blot analysis and inhibition of chymotrypsin.