DNA-mediated transformation in the aquatic filamentous fungus Achlya ambisexualis

Elias Kurian Manavathu, K. Suryanarayana, S. E. Hasnain, W. C. Leung

Research output: Contribution to journalArticle

10 Citations (Scopus)

Abstract

A DNA-mediated transformation system was developed for the aquatic filamentous fungus Achlya ambisexualis using the chimeric plasmid vector pSV2neo2μm. Hyphal colonies resistant to the neomycin analogue G-418 sulphate were regenerated from transformed protoplasts on soft agar. Southern blot analyses of the transformed-cell DNA produced multiple hybridization bands, suggesting integration of vector DNA into the most genome at multiple sites. Northern blot analyses revealed the presence of three APHII-gene-specific transcripts in the transformant, indicating that the G-418-resistant phenotype was due to the expression of the APHII gene. The presence of multiple RNA transcripts of unexpectedly large size suggested tht RNA initiation and/or termination is under the control of regulatory element(s) other than the SV40 promoter. Plasmid DNAs recovered by transformation of Escherichia coli cells with total DNA preparations from the fungal transformants showed considerable DNA rearrangements. However, at least a portion of the plasmid DNA recovered from each of the transformants carried a functional APHII gene, suggesting that the episomal vector DNA may have played a role in maintaining the G-418-resistant phenotype.

Original languageEnglish (US)
Pages (from-to)2019-2028
Number of pages10
JournalJournal of General Microbiology
Volume134
Issue number7
StatePublished - Jan 1 1988

Fingerprint

Achlya
Fungi
DNA
Plasmids
Fungal DNA
RNA
Phenotype
Neomycin
Protoplasts
Gene Rearrangement
Southern Blotting
Northern Blotting
Genes
Agar
Genome
Escherichia coli
Gene Expression

ASJC Scopus subject areas

  • Microbiology

Cite this

Manavathu, E. K., Suryanarayana, K., Hasnain, S. E., & Leung, W. C. (1988). DNA-mediated transformation in the aquatic filamentous fungus Achlya ambisexualis. Journal of General Microbiology, 134(7), 2019-2028.

DNA-mediated transformation in the aquatic filamentous fungus Achlya ambisexualis. / Manavathu, Elias Kurian; Suryanarayana, K.; Hasnain, S. E.; Leung, W. C.

In: Journal of General Microbiology, Vol. 134, No. 7, 01.01.1988, p. 2019-2028.

Research output: Contribution to journalArticle

Manavathu, EK, Suryanarayana, K, Hasnain, SE & Leung, WC 1988, 'DNA-mediated transformation in the aquatic filamentous fungus Achlya ambisexualis', Journal of General Microbiology, vol. 134, no. 7, pp. 2019-2028.
Manavathu EK, Suryanarayana K, Hasnain SE, Leung WC. DNA-mediated transformation in the aquatic filamentous fungus Achlya ambisexualis. Journal of General Microbiology. 1988 Jan 1;134(7):2019-2028.
Manavathu, Elias Kurian ; Suryanarayana, K. ; Hasnain, S. E. ; Leung, W. C. / DNA-mediated transformation in the aquatic filamentous fungus Achlya ambisexualis. In: Journal of General Microbiology. 1988 ; Vol. 134, No. 7. pp. 2019-2028.
@article{a1f2b2a0085a4473b2dfe7eab65397b1,
title = "DNA-mediated transformation in the aquatic filamentous fungus Achlya ambisexualis",
abstract = "A DNA-mediated transformation system was developed for the aquatic filamentous fungus Achlya ambisexualis using the chimeric plasmid vector pSV2neo2μm. Hyphal colonies resistant to the neomycin analogue G-418 sulphate were regenerated from transformed protoplasts on soft agar. Southern blot analyses of the transformed-cell DNA produced multiple hybridization bands, suggesting integration of vector DNA into the most genome at multiple sites. Northern blot analyses revealed the presence of three APHII-gene-specific transcripts in the transformant, indicating that the G-418-resistant phenotype was due to the expression of the APHII gene. The presence of multiple RNA transcripts of unexpectedly large size suggested tht RNA initiation and/or termination is under the control of regulatory element(s) other than the SV40 promoter. Plasmid DNAs recovered by transformation of Escherichia coli cells with total DNA preparations from the fungal transformants showed considerable DNA rearrangements. However, at least a portion of the plasmid DNA recovered from each of the transformants carried a functional APHII gene, suggesting that the episomal vector DNA may have played a role in maintaining the G-418-resistant phenotype.",
author = "Manavathu, {Elias Kurian} and K. Suryanarayana and Hasnain, {S. E.} and Leung, {W. C.}",
year = "1988",
month = "1",
day = "1",
language = "English (US)",
volume = "134",
pages = "2019--2028",
journal = "Microbiology",
issn = "1350-0872",
publisher = "Society for General Microbiology",
number = "7",

}

TY - JOUR

T1 - DNA-mediated transformation in the aquatic filamentous fungus Achlya ambisexualis

AU - Manavathu, Elias Kurian

AU - Suryanarayana, K.

AU - Hasnain, S. E.

AU - Leung, W. C.

PY - 1988/1/1

Y1 - 1988/1/1

N2 - A DNA-mediated transformation system was developed for the aquatic filamentous fungus Achlya ambisexualis using the chimeric plasmid vector pSV2neo2μm. Hyphal colonies resistant to the neomycin analogue G-418 sulphate were regenerated from transformed protoplasts on soft agar. Southern blot analyses of the transformed-cell DNA produced multiple hybridization bands, suggesting integration of vector DNA into the most genome at multiple sites. Northern blot analyses revealed the presence of three APHII-gene-specific transcripts in the transformant, indicating that the G-418-resistant phenotype was due to the expression of the APHII gene. The presence of multiple RNA transcripts of unexpectedly large size suggested tht RNA initiation and/or termination is under the control of regulatory element(s) other than the SV40 promoter. Plasmid DNAs recovered by transformation of Escherichia coli cells with total DNA preparations from the fungal transformants showed considerable DNA rearrangements. However, at least a portion of the plasmid DNA recovered from each of the transformants carried a functional APHII gene, suggesting that the episomal vector DNA may have played a role in maintaining the G-418-resistant phenotype.

AB - A DNA-mediated transformation system was developed for the aquatic filamentous fungus Achlya ambisexualis using the chimeric plasmid vector pSV2neo2μm. Hyphal colonies resistant to the neomycin analogue G-418 sulphate were regenerated from transformed protoplasts on soft agar. Southern blot analyses of the transformed-cell DNA produced multiple hybridization bands, suggesting integration of vector DNA into the most genome at multiple sites. Northern blot analyses revealed the presence of three APHII-gene-specific transcripts in the transformant, indicating that the G-418-resistant phenotype was due to the expression of the APHII gene. The presence of multiple RNA transcripts of unexpectedly large size suggested tht RNA initiation and/or termination is under the control of regulatory element(s) other than the SV40 promoter. Plasmid DNAs recovered by transformation of Escherichia coli cells with total DNA preparations from the fungal transformants showed considerable DNA rearrangements. However, at least a portion of the plasmid DNA recovered from each of the transformants carried a functional APHII gene, suggesting that the episomal vector DNA may have played a role in maintaining the G-418-resistant phenotype.

UR - http://www.scopus.com/inward/record.url?scp=0023764182&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0023764182&partnerID=8YFLogxK

M3 - Article

C2 - 3246595

AN - SCOPUS:0023764182

VL - 134

SP - 2019

EP - 2028

JO - Microbiology

JF - Microbiology

SN - 1350-0872

IS - 7

ER -