Binding of trans-acting protein AngCP to the CCAAT-containing motifs in Aspergillus niger glaA promoter

Xingguo Zhu, Runxiang Qiu, Li Liu, Guomin Tang

Research output: Contribution to journalArticle

Abstract

CCAAT-binding proteins AngCPl and AngCP2 of Aspergillus niger binding to DC (-489 - -414 bp) and PC (-390 - -345 bp) of A. niger glaA gene were respectively purified by 20% - 40% saturated ammonium sulfate, gel filtration, Heparin SepharoseCl-6B chromatography and DNA sequence-specific affinity chromatography. Gel filtration and SDS-PAGE revealed that both AngCPl and AngCP2 were of 120 kD, comprised of two subunits of 34 kD and 50 kD. Western blot showed that the 34 kD subunits of both AngCPl and AngCP2 cross-reacted specifically with the anti-AngHAPC antiserum. Further electrophoretic mobility shift assay identified that AngCPl and AngCP2 were the same protein, designated AngCP. Southwestern blot showed that the affinity of the 34 kD sub-unit to DC was stronger than that of the 50 kD subunit to PC. These results suggested that interaction between AngCP, DC and PC plays an important role in the regulation of transcription of Aspergillus niger glaA gene.

Original languageEnglish (US)
Pages (from-to)338-343
Number of pages6
JournalProgress in Natural Science
Volume14
Issue number4
DOIs
StatePublished - Jan 1 2004
Externally publishedYes

Fingerprint

Aspergillus
Gels
Genes
Proteins
Affinity chromatography
Electrophoretic mobility
DNA sequences
Ammonium Sulfate
Transcription
Chromatography
Heparin
Immune Sera
Assays
Carrier Proteins
Sulfates

Keywords

  • Aspergillus niger glucoamylase
  • CCAAT-binding protein
  • DNA-protein interaction
  • Protein purification

ASJC Scopus subject areas

  • Materials Science(all)

Cite this

Binding of trans-acting protein AngCP to the CCAAT-containing motifs in Aspergillus niger glaA promoter. / Zhu, Xingguo; Qiu, Runxiang; Liu, Li; Tang, Guomin.

In: Progress in Natural Science, Vol. 14, No. 4, 01.01.2004, p. 338-343.

Research output: Contribution to journalArticle

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AU - Tang, Guomin

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AB - CCAAT-binding proteins AngCPl and AngCP2 of Aspergillus niger binding to DC (-489 - -414 bp) and PC (-390 - -345 bp) of A. niger glaA gene were respectively purified by 20% - 40% saturated ammonium sulfate, gel filtration, Heparin SepharoseCl-6B chromatography and DNA sequence-specific affinity chromatography. Gel filtration and SDS-PAGE revealed that both AngCPl and AngCP2 were of 120 kD, comprised of two subunits of 34 kD and 50 kD. Western blot showed that the 34 kD subunits of both AngCPl and AngCP2 cross-reacted specifically with the anti-AngHAPC antiserum. Further electrophoretic mobility shift assay identified that AngCPl and AngCP2 were the same protein, designated AngCP. Southwestern blot showed that the affinity of the 34 kD sub-unit to DC was stronger than that of the 50 kD subunit to PC. These results suggested that interaction between AngCP, DC and PC plays an important role in the regulation of transcription of Aspergillus niger glaA gene.

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