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 language | English (US) |
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Pages (from-to) | 53-59 |
Number of pages | 7 |
Journal | Gene |
Volume | 57 |
Issue number | 1 |
DOIs | |
State | Published - 1987 |
Externally published | Yes |
Keywords
- DNA transformation
- Recombinant DNA
- Southern blot
- metallothionein promoter
- plasmid vector
- thymidine incorporation
ASJC Scopus subject areas
- Genetics