Normalization of transfection efficiency using the β-lactamase gene of the pGL3 luciferase vector in primary anterior pituitary cells

Seong W. Kang, Seungkwon You, Eric A. Wong, Mohamed E. El Halawani

Research output: Contribution to journalArticle

2 Citations (Scopus)

Abstract

The β-galactosidase reporter gene is commonly used as a control for transfection efficiency in the promoter reporter assay system. While investigating vasoactive intestinal peptide response elements in the promoter of the prolactin gene, we found that primary pituitary cells from turkey hens highly expressed endogenous β-galactosidase. Therefore, we developed a new protocol for determining transfection efficiency using the β-lactamase gene, which is present on many expression vectors. Transcript levels of β-lactamase were measured by RT-PCR after transfection of different amounts of the pGL3-basic and pGL3-control vectors. A high correlation was observed between the amount of plasmid transfected and β-lactamase mRNA levels. Although no eukaryotic promoter was present, there was apparently leaky expression of the β-lactamase gene. Expression of β-lactamase was independent of expression from the simian virus 40 or turkey prolactin promoters cloned upstream of the luciferase gene.

Original languageEnglish
Pages (from-to)326-330
Number of pages5
JournalBioTechniques
Volume33
Issue number2
Publication statusPublished - 2002 Jan 1
Externally publishedYes

Fingerprint

Luciferases
Galactosidases
Transfection
Genes
Prolactin
Simian virus 40
Vasoactive Intestinal Peptide
Response Elements
Reporter Genes
Plasmids
Gene Expression
Polymerase Chain Reaction
Messenger RNA
Viruses
Assays

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)
  • Clinical Biochemistry

Cite this

Normalization of transfection efficiency using the β-lactamase gene of the pGL3 luciferase vector in primary anterior pituitary cells. / Kang, Seong W.; You, Seungkwon; Wong, Eric A.; El Halawani, Mohamed E.

In: BioTechniques, Vol. 33, No. 2, 01.01.2002, p. 326-330.

Research output: Contribution to journalArticle

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