Expression of herpes simplex virus thymidine kinase gene in aquatic filamentous fungus achlya ambisexualis

Elias K. Manavathu, Kalachar Suryanarayana, Seyed E. Hasnain, Maria Leung, Fai Lau Yun Fai Lau, Wai Choi Leung

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2 Scopus citations

Abstract

We have constructed a plasmid vector pSV2neo-MKαG in which the structural tk gene for Herpes simplex virus thymidine kinase (HSV-TK) was placed downstream from the metallothionein-I promoter. The vector also contained the selection marker aminoglycoside 3′-phosphotransferase (Kmr). This vector was able to transform the filamentous fungus Achlya ambisexualis and G-418-resistant colonies were obtained. Southern blot analyses revealed that multiple bands hybridizing to the HSV tk gene probe were present in the genomic DNA of the transformants. Upon analysis by gel electrophoresis, one of the transformants exhibited TK activity bearing electrophoretic mobility similar to that of the HSV-TK. An increase of approx. 40% of [3H]thymidine uptake and incorporation into cellular DNA was also observed in this transformant. This study suggested that the HSV tk gene can be expressed in the fungus A. ambisexualis that can be considered as a candidate host cell for further gene-expression studies.

Original languageEnglish (US)
Pages (from-to)53-59
Number of pages7
JournalGene
Volume57
Issue number1
DOIs
StatePublished - 1987

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Keywords

  • DNA transformation
  • Recombinant DNA
  • Southern blot
  • metallothionein promoter
  • plasmid vector
  • thymidine incorporation

ASJC Scopus subject areas

  • Genetics

Cite this

Manavathu, E. K., Suryanarayana, K., Hasnain, S. E., Leung, M., Yun Fai Lau, F. L., & Leung, W. C. (1987). Expression of herpes simplex virus thymidine kinase gene in aquatic filamentous fungus achlya ambisexualis. Gene, 57(1), 53-59. https://doi.org/10.1016/0378-1119(87)90176-4