[Construction of cell lines producing high-titer recombinant retroviruses of message and antisense cDNA of human glucocorticoid receptor beta]

Zhong Nan Da Xue Xue Bao Yi Xue Ban. 2004 Oct;29(5):540-3.
[Article in Chinese]

Abstract

Objective: To construct cell lines producing high-titer recombinant retroviruses of human glucocorticoid receptor beta (hGRbeta).

Methods: The recombinant retrorinal vectors of message and antisense cDNA of hGRbeta were transfected into PA317 cells by liposomes, respectively, and cell colonies were selected with G418. The titers of viruses from the 2 cell culture supernatants were determined by colony numbers produced in NIH3T3 cells.

Results: The presence of aimed RNAs in supernatant of the G418-resistant cell cultures were detected by RT-PCR and subsequent sequence analysis. The titers of retroviruses were 2. 0 x 10(5) and 1.8 x 10(5), each for recombinant message and antisense hGRbeta cDNA.

Conclusion: We demonstrated here the obtaining of transfect PA317 cells that produce recombinant retroviruses containing message and antisense RNAs of hGRbeta with highly infective efficiencies.

Publication types

  • English Abstract
  • Research Support, Non-U.S. Gov't

MeSH terms

  • 3T3 Cells
  • Animals
  • Cell Line
  • Cloning, Molecular
  • DNA, Antisense / genetics*
  • DNA, Complementary / genetics
  • Genetic Vectors
  • Humans
  • Mice
  • Receptors, Glucocorticoid / biosynthesis*
  • Receptors, Glucocorticoid / genetics
  • Recombinant Proteins / biosynthesis
  • Recombinant Proteins / genetics
  • Retroviridae / genetics*
  • Transfection

Substances

  • DNA, Antisense
  • DNA, Complementary
  • Receptors, Glucocorticoid
  • Recombinant Proteins
  • glucocorticoid receptor beta