Expression cloning of a mammalian amino acid transporter or modifier by complementation of a yeast transport mutant

Lin, Guorong; Cai, Xiao; Johnstone, Rose M.

doi:10.1002/(sici)1097-4652(199712)173:3<351::aid-jcp7>3.0.co;2-m

Cited by 2 publications

(2 citation statements)

References 30 publications

(6 reference statements)

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“…However, the characteristics and splicing mechanisms of yeast introns are different from those of filamentous fungi (Gurr et al ., 1987). In yeast, no requirement for the presence of introns has been observed when genes from plants or vertebrates were expressed (Hiraiwa et al ., 1997; Lin et al ., 1997). In contrast to introns in genes from animals, S. cerevisiae and plants, introns of filamentous fungi are characteristically short (on average less than 100 bp).…”

Section: Discussionmentioning

confidence: 99%

Introns are necessary for mRNA accumulation in Schizophyllum commune

Lugones

Scholtmeijer

Klootwijk

et al. 1999

Molecular Microbiology

101

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The cDNA coding sequence of the Agaricus bisporus hydrophobin gene ABH1 under the regulation sequences of the Schizophyllum commune SC3 hydrophobin gene gave no expression in S. commune. In contrast, the genomic coding sequence (containing three introns) produced high levels of ABH1 mRNA when transformed to S. commune in the same configuration. Apparently, introns were needed for the accumulation of mRNAs from the ABH1 gene. When the effect of intron deletion on expression of the homologous genes SC3 and SC6 was examined, it was observed that only the genomic coding sequences were expressed in S. commune. Run-on analysis with nuclei harbouring intron-containing and intronless SC6 showed that this effect did not occur at the level of transcription initiation: genomic and cDNA sequences were equally active in this respect. When a 50 bp artificial intron containing the consensus splice and branch sites of S. commune introns, in addition to random-generated sequences, was introduced in the right orientation into the intronless SC3 transcriptional unit, accumulation of SC3 mRNA was restored. By polymerase chain reaction amplification, no unspliced SC3 mRNA species could be detected. Furthermore, the addition of an intron into the transcriptional unit of the gene for green fluorescent protein (GFP) effected clear fluorescence of the transgenic hyphae. Apparently, splicing is required for the normal processing of primary transcripts in S. commune.

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Section: Discussionmentioning

confidence: 99%

Introns are necessary for mRNA accumulation in Schizophyllum commune

Lugones

Scholtmeijer

Klootwijk

et al. 1999

Molecular Microbiology

101

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“…Xu and Gong (2003) showed that introns are required for AFP gene (a gene encoding antifungal protein in A. giganteus) expression in T. atroviride transformants, as demonstrated by the level of mRNA and confirmed by analysis of AFP synthesis. In contrast, in the ascomycetous yeast S. cerevisiae only 2 to 5% of the genes contain introns and no obligation for introns has been observed for vertebrate and plant genes expression (Hiraiwa et al, 1997;Lin et al, 1997). However, less is even known about intron-dependent mRNA accumulation in fungi and further researches should be accomplished.…”

Section: The Use Of Native or Artificial Intron-containing Genes In Fmentioning

confidence: 99%

Genetic manipulation of fungal strains for the improvement of heterologous genes expression (a mini-review)

Koushki¹,

Hamid²,

Farsi³

2011

Afr. J. Biotechnol.

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Fungi are eukaryotic microorganisms that influence our everyday lives in areas as diverse as agriculture, medicine and basic science. With the advent of molecular biology, it has been attempted to improve the expression level of heterologous genes, which encode favorable traits in fungal strains. The expression of heterologous genes can be limited in transcription, post-transcription, translation and post-translation levels. Several genetic strategies have been developed to reduce the expression constrains and to enhance genes functionality. Among these strategies can be pointed to the introduction of multicopies of the desired gene, change of AT-rich sequences, gene fusion with a wellexpressed gene, the use of strong promoters and signal sequences, optimization of codon usage, the construction and use of protease-deficient and chaperones/foldases-overproduced strains and the use of native or artificial intron-containing genes. These strategies have often resulted in the expected increase in the expression of heterologus genes. With the isolation of a large number of genes encoding desired traits and the availability of a large collection of wild isolates, the improvement of strains with a better functional performance would be possible.

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Expression cloning of a mammalian amino acid transporter or modifier by complementation of a yeast transport mutant

Cited by 2 publications

References 30 publications

Introns are necessary for mRNA accumulation in Schizophyllum commune

Introns are necessary for mRNA accumulation in Schizophyllum commune

Genetic manipulation of fungal strains for the improvement of heterologous genes expression (a mini-review)

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