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

Eur J Biochem. 1986 Oct 1;160(1):61-7. doi: 10.1111/j.1432-1033.1986.tb09940.x.

Abstract

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.

MeSH terms

  • Amino Acid Sequence
  • Base Sequence
  • Cervix Uteri / analysis*
  • Cloning, Molecular*
  • DNA / chemical synthesis
  • Escherichia coli / genetics
  • Female
  • Humans
  • Nucleic Acid Hybridization
  • Oligonucleotides / chemical synthesis
  • Plasmids
  • Protease Inhibitors / genetics*
  • Proteinase Inhibitory Proteins, Secretory
  • Proteins*
  • RNA, Messenger / isolation & purification
  • Transformation, Genetic

Substances

  • Oligonucleotides
  • Protease Inhibitors
  • Proteinase Inhibitory Proteins, Secretory
  • Proteins
  • RNA, Messenger
  • DNA

Associated data

  • GENBANK/X04470