Identification and functional analysis of beta-tubulin genes by site specific integrative transformation in Aspergillus nidulans.

G S May, J Gambino, J A Weatherbee, N R Morris
Author Information

Abstract

We have cloned two different beta-tubulin sequences from the filamentous fungus Aspergillus nidulans. Each was used in the construction of transforming plasmids that carry the pyr4 gene of Neurospora crassa. We used these plasmids to transform a pyrG-strain of Aspergillus to uridine prototrophy. Both plasmids were shown to integrate site specifically into the homologous chromosomal sequences. We then used transformant strains in genetic crosses to demonstrate that one of the cloned beta-tubulin sequences was the benA beta-tubulin gene, which codes for the beta 1-and beta 2-tubulins. The other cloned beta-tubulin sequence was shown to be the structural gene for beta 3-tubulin by gene disruption and to participate in conidial development. This is the first report of a gene disruption by site specific, integrative recombination in Aspergillus nidulans.

References

  1. J Mol Biol. 1970 Oct 14;53(1):159-62 [PMID: 4922220]
  2. J Cell Biol. 1985 Sep;101(3):706-11 [PMID: 3897246]
  3. Mol Gen Genet. 1975 Jun 19;138(3):243-55 [PMID: 1102932]
  4. J Mol Biol. 1975 Nov 5;98(3):503-17 [PMID: 1195397]
  5. J Mol Biol. 1977 Jun 15;113(1):237-51 [PMID: 881736]
  6. Cell. 1978 Oct;15(2):639-47 [PMID: 363278]
  7. Cell. 1979 Feb;16(2):437-42 [PMID: 378391]
  8. Cell. 1980 Dec;22(3):845-54 [PMID: 6780200]
  9. Nature. 1981 Feb 19;289(5799):650-5 [PMID: 7464932]
  10. Cell. 1981 Jun;24(3):837-45 [PMID: 7018696]
  11. Anal Biochem. 1981 Jun;114(1):193-7 [PMID: 6269464]
  12. Proc Natl Acad Sci U S A. 1981 Aug;78(8):4877-81 [PMID: 6946435]
  13. Biochim Biophys Acta. 1981 Dec 28;656(2):195-205 [PMID: 6172152]
  14. Gene. 1982 Oct;19(3):259-68 [PMID: 6295879]
  15. Biochem Biophys Res Commun. 1983 Apr 15;112(1):284-9 [PMID: 6220717]
  16. Proc Natl Acad Sci U S A. 1982 Oct;79(19):5976-80 [PMID: 6764535]
  17. Nature. 1983 Sep 29-Oct 5;305(5933):391-7 [PMID: 6353245]
  18. Gene. 1983 Dec;26(1):101-6 [PMID: 6323249]
  19. Gene. 1983 Dec;26(2-3):205-21 [PMID: 6368319]
  20. J Cell Biol. 1984 Jan;98(1):1-7 [PMID: 6707079]
  21. Proc Natl Acad Sci U S A. 1984 Mar;81(5):1470-4 [PMID: 6324193]
  22. Cell. 1983 May;33(1):211-9 [PMID: 6380751]
  23. Genetics. 1984 Sep;108(1):123-41 [PMID: 6479584]
  24. Genetics. 1984 Sep;108(1):143-64 [PMID: 6090267]
  25. J Biol Chem. 1984 Dec 25;259(24):15452-9 [PMID: 6210290]
  26. Biochim Biophys Acta. 1973 Apr 11;299(4):516-20 [PMID: 4575181]

MeSH Term

Aspergillus nidulans
Benomyl
Cloning, Molecular
Genes, Fungal
Isoelectric Point
Molecular Weight
Mutation
Plasmids
Recombination, Genetic
Transformation, Genetic
Tubulin

Chemicals

Tubulin
Benomyl

Word Cloud

Created with Highcharts 10.0.0beta-tubulingeneAspergillusclonedsequencesnidulansusedplasmidssitebetashowndisruptionspecificintegrativetwodifferentfilamentousfungusconstructiontransformingcarrypyr4NeurosporacrassatransformpyrG-strainuridineprototrophyintegratespecificallyhomologouschromosomaltransformantstrainsgeneticcrossesdemonstrateonebenAcodes1-and2-tubulinssequencestructural3-tubulinparticipateconidialdevelopmentfirstreportrecombinationIdentificationfunctionalanalysisgenestransformation

Similar Articles

Cited By