Molecular characterization of a stably transformed Bombyx mon cell line: Identification of alternative transcriptional initiation sites of the A3 cytoplasmic actin gene

K. Fatyol, K. Illes, T. Praznovszky, W. H R Langridge, G. Hadlaczky, A. A. Szalay

Research output: Contribution to journalArticle

4 Citations (Scopus)

Abstract

We have isolated a stably transformed Bombyx mori cell line containing a novel selectable marker gene, puromycin N-acetyl transferase, under control of transcriptional regulatory signals from the A3 cytoplasmic actin gene. By using this cell line we have identified alternative transcriptional initiation sites for the A3 actin gene. One of these start sites is located ∼35 bp upstream from the previously determined transcription initiation site. The two mRNA start sites are utilized with a similar efficiencies in the BmN cell line. In addition, we detected transcripts that initiated in the first intron of the A3 actin gene. These transcripts may be synthesised under control of an alternative promoter. The stably transformed B. mori cell line used in this study was also extensively characterized. Integration of the plasmid molecules into the host genome was demonstrated by Southern and in situ hybridization analyses. Establishment and characterization of stably transformed insect cell lines, like the one described here, represents an important step in the development of non-lytic insect expression systems.

Original languageEnglish
Pages (from-to)1-8
Number of pages8
JournalMolecular Genetics and Genomics
Volume255
Issue number5
Publication statusPublished - Oct 2 1998

Fingerprint

Bombyx
Actins
Cell Line
Genes
Insects
Puromycin
Transformed Cell Line
Transcription Initiation Site
Transferases
Introns
In Situ Hybridization
Plasmids
Genome
Messenger RNA

Keywords

  • 5′ RACE
  • A3 cytoplasmic actin
  • Puromycin N-acetyl transferase
  • Silkworm
  • Stably transformed cell line

ASJC Scopus subject areas

  • Genetics

Cite this

Molecular characterization of a stably transformed Bombyx mon cell line : Identification of alternative transcriptional initiation sites of the A3 cytoplasmic actin gene. / Fatyol, K.; Illes, K.; Praznovszky, T.; Langridge, W. H R; Hadlaczky, G.; Szalay, A. A.

In: Molecular Genetics and Genomics, Vol. 255, No. 5, 02.10.1998, p. 1-8.

Research output: Contribution to journalArticle

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AU - Hadlaczky, G.

AU - Szalay, A. A.

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AB - We have isolated a stably transformed Bombyx mori cell line containing a novel selectable marker gene, puromycin N-acetyl transferase, under control of transcriptional regulatory signals from the A3 cytoplasmic actin gene. By using this cell line we have identified alternative transcriptional initiation sites for the A3 actin gene. One of these start sites is located ∼35 bp upstream from the previously determined transcription initiation site. The two mRNA start sites are utilized with a similar efficiencies in the BmN cell line. In addition, we detected transcripts that initiated in the first intron of the A3 actin gene. These transcripts may be synthesised under control of an alternative promoter. The stably transformed B. mori cell line used in this study was also extensively characterized. Integration of the plasmid molecules into the host genome was demonstrated by Southern and in situ hybridization analyses. Establishment and characterization of stably transformed insect cell lines, like the one described here, represents an important step in the development of non-lytic insect expression systems.

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