CDNA CLONING AND EXPRESSION OF A TALAROMYCES-EMERSONII AMYLASE ENCODING GENETIC DETERMINANT IN ESCHERICHIA-COLI

L BUNNI, DC COLEMAN, L MCHALE, TJ HACKETT, AP MCHALE

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Abstract

Intact mRNA has been isolated from the thermophilic fungus Talaromyces emersonii following grow on starch containing media. This has been used as template to synthesise cDNA. The cDNA was cloned into the Escherichia coli expression vector system, pUC18 and this was used to transform E. coli. Transformed colonies were screened for production of amylase activity and a number of positive recombinants were detected. One of those was found to contain a plasmid named pMH1, which harboured a 1.2 kb insert. Sub-cloning experiments verified that the amylase phenotype was encoded for by this fragment. The fragment was characterised using restriction enzyme cleavage site analysis. The origin of the insert was verified using both Northern and Southern blotting hybridisation analysis of T. emersonii RNA and DNA respectively.
LanguageEnglish
Pages1109-1114
JournalBiotechnology Letters
Volume14
Issue number12
DOIs
Publication statusPublished - Dec 1992

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amylases
molecular cloning
thermophilic fungi
Escherichia coli
Northern blotting
Southern blotting
plasmids
hybridization
starch
RNA
phenotype
DNA
enzymes
Talaromyces emersonii

Cite this

BUNNI, L ; COLEMAN, DC ; MCHALE, L ; HACKETT, TJ ; MCHALE, AP. / CDNA CLONING AND EXPRESSION OF A TALAROMYCES-EMERSONII AMYLASE ENCODING GENETIC DETERMINANT IN ESCHERICHIA-COLI. In: Biotechnology Letters. 1992 ; Vol. 14, No. 12. pp. 1109-1114.
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abstract = "Intact mRNA has been isolated from the thermophilic fungus Talaromyces emersonii following grow on starch containing media. This has been used as template to synthesise cDNA. The cDNA was cloned into the Escherichia coli expression vector system, pUC18 and this was used to transform E. coli. Transformed colonies were screened for production of amylase activity and a number of positive recombinants were detected. One of those was found to contain a plasmid named pMH1, which harboured a 1.2 kb insert. Sub-cloning experiments verified that the amylase phenotype was encoded for by this fragment. The fragment was characterised using restriction enzyme cleavage site analysis. The origin of the insert was verified using both Northern and Southern blotting hybridisation analysis of T. emersonii RNA and DNA respectively.",
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CDNA CLONING AND EXPRESSION OF A TALAROMYCES-EMERSONII AMYLASE ENCODING GENETIC DETERMINANT IN ESCHERICHIA-COLI. / BUNNI, L; COLEMAN, DC; MCHALE, L; HACKETT, TJ; MCHALE, AP.

In: Biotechnology Letters, Vol. 14, No. 12, 12.1992, p. 1109-1114.

Research output: Contribution to journalArticle

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