Molecular Cloning, Lentiviral Transduction, and Expression of Recombinant ADAMTSL2 and ADAMTSL4

Methods Mol Biol. 2020:2043:137-155. doi: 10.1007/978-1-4939-9698-8_12.

Abstract

Lentiviral systems have proven advantageous in the delivery and long-term integration of gene sequences into the genome of several cell types in vitro, in vivo, as well as in clinical trials. Here we detail the protocols involved in the molecular cloning of ADAMTSL2 and ADAMTSL4 into the human immunodeficiency virus (HIV)-derived pCDH lentiviral system. We also describe the lentiviral transduction of ADAMTSL2 and ADAMTSL4 into mammalian HEK293-EBNA cells to create stable cell lines, as well as their recombinant expression.

Keywords: HEK293-EBNA; Lentiviral system; Lentiviral transduction; Mammalian; Molecular cloning; Recombinant expression; Stable cell lines.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • ADAMTS Proteins / genetics*
  • ADAMTS Proteins / metabolism
  • Cloning, Molecular / methods*
  • Genetic Vectors
  • HEK293 Cells
  • Humans
  • Lentivirus / genetics
  • Recombinant Proteins / metabolism
  • Transduction, Genetic / methods*

Substances

  • ADAMTSL4 protein, human
  • Recombinant Proteins
  • ADAMTS Proteins
  • ADAMTSL2 protein, human