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Alcohol dehydrogenase 5 of Helicoverpa armigera interacts with the CYP6B6 promoter in response to 2-tridecanone

Insect Science 2020年第5期27卷 1053-1066页

作者： Jie Zhao Qian Wei Xin-Rong Gu Su-Wei Ren Xiao-Ning Liu Xinjiang Key Laboratory of Biological Resources and Genetic Engineering College of Life Sciences and TechnologyXinjiang UniversityUrumqiXinjiangChina Key Laboratory of Oasis Agricultural Pest Management and Plant Protection Resources Utilization College of AgricultureShihezi UniversityShiheziXinjiangChina

Alcohol dehydrogenase 5(ADH5)is a member of medium-chain dehydro-genase/reductase family and takes part in cellular formaldehyde and S-nitrosoglutathione metabolic network.2-tridecanone(2-TD)is a toxic compound in many Solanaceae crops to defend against a variety of herbivory *** the broader context of insect development and pest control strategies,this study investigates how a new ADHS from Helicoverpa armigera(HaADH5)regulates the expression of CYP6B6,a gene involved in molting and metamorphosis,in response to 2-TD *** of the HaADH5 complementary dna sequence revealed that its 1002 bp open reading frame encodes 334 amino acids with a predicted molecular weight of 36.5 ***5 protein was purified in the Es-cherichia coli Transetta(pET32a-HaADH5)strain using a prokaryotic expression *** ability of HaADH5 protein to interact with the 2-TD responsive region within the promoter of CYP6B6 was confirmed by an in vitro electrophoretic mobility shift assay and transcription activity validation in ***,the expression levels of both HaADH5 and CYP6B6 were found to be significantly decreased in the midgut of 6th instar larvae after 48 h of treatment with 10 mg/g 2-TD artificial *** results indicate that upon 2-TD treatment of cotton bollworm,HaADHS regulates the expression of CYP6B6 by interacting with its *** HaADH5 regulation of CYP6B6 expression may con-tribute to the larval xenobiotic detoxification,molting and metamorphosis,HaADH5 is *** target for controlling the growth and development of cotton bollworm.

关键词： alcohol dehydrogenase dna-protein interaction Helicoverpa armigera regulatory factor 2-tridecanone

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

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Progress in Natural Science:Materials International 2004年第4期 52-57页

作者： ZHU Xingguo, QIU Runxiang, LIU Li and TANG Guomin(Institute of Microbiology, Chinese Academy of Sciences, Beijing 100080, China)

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.

关键词： Aspergillus niger glucoamylase CCAAT-binding protein dna-protein interaction protein purification.

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2-（2-azidoethyl）-2,5-di-（4,5-Dimethoxy-2-nitro-benzyloxy）-2,5-dihydr ofuran as a potential photoaffinity crosslinking reagent

2-（2-azidoethyl）-2,5-di-（4,5-Dimethoxy-2-nitro-benzyloxy...

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第十届全国化学生物学学术会议

作者： Jiahui Li Mengdi Shang Xiaoping Zhou Zhen Xi Chuanzheng Zhou State Key Laboratory of Elemento-Organic Chemistry and Department of Chemical Biology College of ChemistryNankai University

Photoaffinity crosslinking is widely used in biomolecules interaction studies. A few photoreactive groups, such as azide, diazirine and benzophenone, have introduced to fulfill such a purpose. However, all of these groups suffer from low cross-linking efficiency as a result of either low reactivity or photodecomposition. We design and synthesized 2-（2-azidoethyl）-2,5-di-（4,5-Dimethoxy-2-nitro-benzyloxy）-2,5-dihydrofuran as a new photoaffinity crosslinking reagent. The azid function on this molecule allow it to be readily anchored to alkyne modified biomolecules. Photo irradiation of the molecule at 350 nm furnish cis-enedione, which is quite stable but crosslink with primary amines quickly and quantitatively. The unique property make this molecule prominent in mapping out protein residues involved in protein–dna interactions and to trapping/identifying proteins that interact with dna.

关键词： Photoaffinity crosslinking dna-protein interaction target protein

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