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

作　　者：ZHUXingguo QIURunxiang LIULi TANGGuomin 

机构地区：[1]InstituteofMicrobiology,ChineseAcademyofSciences,Beijing100080,China

出　　处：《Progress in Natural Science:Materials International》2004年第4期338-343,共6页自然科学进展·国际材料（英文版）

基　　金：Supported by the National Natural Science Foundation of China (No. 30170014)

摘　　要：CCAAT-binding proteins AngCP1 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, HeparinSepharoseCl-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.CCAAT-binding proteins AngCP1 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-speciflc affinity chromatography. Gel filtration and SDS-PAGE revealed that both AngCP1 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 AngCP1 and AngCP2 cross-reacted specifically with the anti-AngHAPC antiserum. Further electrophoretic mobility shift assay identified that AngCP1 and AngCP2 were the same protein, designated AngCP. Southwestern blot showed that the affinity of the 34 kD subunit 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.

关 键 词：Aspergillus niger glucoamylase CCAAT-binding protein DNA-protein interaction protein purification. 

分 类 号：Q949.327.1[生物学—植物学]