Gene synthesis and expression in E. coli for pump, a human matrix metalloproteinase

Qizhuang Ye, Linda L. Johnson, Vijaykumar Baragi

Research output: Contribution to journalArticle

34 Citations (Scopus)

Abstract

The gene for PUMP (putative metalloproteinase), a human matrix metalloproteinase, was synthesized by a PCR-based method. The DNA fragment of 546 bases containing the PUMP gene was generated by overlap extension of six long oligonucleotides (length ranging from 101 to 116 bases) and subsequent amplification by two short terminal oligonucleotide primers (length from 20 to 48 bases) in one pot without using restriction and ligation enzymes. The synthetic gene was cloned into a T7 expression vector in two ways to express PUMP as a non-fusion protein. Both constructs showed high level expression in E. coli.

Original languageEnglish (US)
Pages (from-to)143-149
Number of pages7
JournalBiochemical and Biophysical Research Communications
Volume186
Issue number1
DOIs
StatePublished - Jul 15 1992
Externally publishedYes

Fingerprint

Metalloproteases
Matrix Metalloproteinases
Escherichia coli
Genes
Pumps
Gene Expression
Synthetic Genes
DNA Primers
Oligonucleotides
Ligation
Amplification
Polymerase Chain Reaction
DNA
Enzymes
Proteins

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Molecular Biology

Cite this

Gene synthesis and expression in E. coli for pump, a human matrix metalloproteinase. / Ye, Qizhuang; Johnson, Linda L.; Baragi, Vijaykumar.

In: Biochemical and Biophysical Research Communications, Vol. 186, No. 1, 15.07.1992, p. 143-149.

Research output: Contribution to journalArticle

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