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.
D000595:Amino Acid Sequence; D001483:Base Sequence; D002584:Cervix Uteri / Q000032:analysis*; D003001:Cloning, Molecular; D004247:DNA / Q000138:chemical synthesis; D004926:Escherichia coli / Q000235:genetics; D005260:Female; D006801:Humans; D009693:Nucleic Acid Hybridization; D009841:Oligonucleotides / Q000138:chemical synthesis; D010957:Plasmids; D011480:Protease Inhibitors / Q000235:genetics*; D053491:Proteinase Inhibitory Proteins, Secretory; D011506:Proteins; D012333:RNA, Messenger / Q000302:isolation & purification; D014170:Transformation, Genetic
D009841:Oligonucleotides; D011480:Protease Inhibitors; D053491:Proteinase Inhibitory Proteins, Secretory; D011506:Proteins; D012333:RNA, Messenger; D004247:DNA