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Appl Environ Microbiol. 1999 May;65(5):2250-2. doi: 10.1128/AEM.65.5.2250-2252.1999.
2
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An abundant hydrophobin (ABH1) forms hydrophobic rodlet layers in Agaricus bisporus fruiting bodies.一种丰富的疏水蛋白（ABH1）在双孢蘑菇子实体中形成疏水小杆层。
Microbiology (Reading). 1996 May;142 ( Pt 5):1321-1329. doi: 10.1099/13500872-142-5-1321.
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Identification of genes associated with morphology in Aspergillus niger by using suppression subtractive hybridization.利用抑制性消减杂交技术鉴定黑曲霉中与形态相关的基因。
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A novel expression system for intracellular production and purification of recombinant affinity-tagged proteins in Aspergillus niger.一种新型表达系统，用于在黑曲霉细胞内生产和纯化重组亲和标签蛋白。
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引用本文的文献

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Production of the Aspergillus aculeatus endo-1,4-beta-mannanase in A. niger.棘孢曲霉内切-1,4-β-甘露聚糖酶在黑曲霉中的产生。
J Ind Microbiol Biotechnol. 2009 Apr;36(4):611-7. doi: 10.1007/s10295-009-0551-x. Epub 2009 Mar 10.

本文引用的文献

1
Secretion of active human mucus proteinase inhibitor by Aspergillus niger after KEX2-like processing of a glucoamylase-inhibitor fusion protein.黑曲霉对葡糖淀粉酶抑制剂融合蛋白进行类KEX2加工后分泌活性人黏液蛋白酶抑制剂
J Biotechnol. 1996 Dec 10;52(2):97-106. doi: 10.1016/s0168-1656(96)01634-3.
2
Hydrophobins: proteins that change the nature of the fungal surface.疏水蛋白：改变真菌表面性质的蛋白质。
Adv Microb Physiol. 1997;38:1-45. doi: 10.1016/s0065-2911(08)60154-x.
3
An abundant hydrophobin (ABH1) forms hydrophobic rodlet layers in Agaricus bisporus fruiting bodies.一种丰富的疏水蛋白（ABH1）在双孢蘑菇子实体中形成疏水小杆层。
Microbiology (Reading). 1996 May;142 ( Pt 5):1321-1329. doi: 10.1099/13500872-142-5-1321.
4
The Agaricus bisporus hypA gene encodes a hydrophobin and specifically accumulates in peel tissue of mushroom caps during fruit body development.双孢蘑菇hypA基因编码一种疏水蛋白，在子实体发育过程中特异性地积累在蘑菇菌盖的表皮组织中。
J Mol Biol. 1996 Apr 19;257(5):1008-18. doi: 10.1006/jmbi.1996.0219.
5
Ultrasensitive stain for proteins in polyacrylamide gels shows regional variation in cerebrospinal fluid proteins.聚丙烯酰胺凝胶中蛋白质的超敏染色显示脑脊液蛋白质存在区域差异。
Science. 1981 Mar 27;211(4489):1437-8. doi: 10.1126/science.6162199.
6
Development of a homologous transformation system for Aspergillus niger based on the pyrG gene.基于pyrG基因的黑曲霉同源转化系统的开发。
Mol Gen Genet. 1987 Jan;206(1):71-5. doi: 10.1007/BF00326538.
7
Transformation of Aspergillus based on the hygromycin B resistance marker from Escherichia coli.基于来自大肠杆菌的潮霉素B抗性标记对曲霉进行转化。
Gene. 1987;56(1):117-24. doi: 10.1016/0378-1119(87)90164-8.
8
Localization of growth and secretion of proteins in Aspergillus niger.
J Gen Microbiol. 1991 Aug;137(8):2017-23. doi: 10.1099/00221287-137-8-2017.
9
Visualization of proteases within a complex sample following their selective retention on immobilized bacitracin, a peptide antibiotic.在蛋白酶选择性保留于固定化杆菌肽（一种肽类抗生素）后，对复杂样品中的蛋白酶进行可视化分析。
Anal Biochem. 1991 Nov 1;198(2):385-90. doi: 10.1016/0003-2697(91)90444-x.
10
Isolation and characterization of mutants of Aspergillus niger deficient in extracellular proteases.黑曲霉胞外蛋白酶缺陷型突变体的分离与鉴定
Mol Gen Genet. 1992 Aug;234(2):332-6. doi: 10.1007/BF00283855.

一种用于评估丝状真菌中异源蛋白产量的夹心培养技术。

A sandwiched-culture technique for evaluation of heterologous protein production in a filamentous fungus.

作者信息

Asgeirsdóttir S A, Scholtmeijer K, Wessels J G

机构信息

Molecular Plant Biology Laboratory, Biomolecular Sciences and Biotechnology Institute, University of Groningen, 9751 NN Haren, The Netherlands.

出版信息

Appl Environ Microbiol. 1999 May;65(5):2250-2. doi: 10.1128/AEM.65.5.2250-2252.1999.

DOI:10.1128/AEM.65.5.2250-2252.1999
PMID:10224030
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC91327/
Abstract

Aspergillus niger is known for its efficient excretion machinery. However, problems have often arisen in obtaining high amounts of heterologous proteins in the culture medium. Here we present a quick method using sandwiched colonies to evaluate transgenic strains for secretion of heterologous proteins. Expressing the ABH1 hydrophobin of Agaricus bisporus in A. niger, we showed that low production levels of the heterologous protein are probably due to extracellular proteolytic degradation of the protein.

摘要

黑曲霉以其高效的分泌机制而闻名。然而，在培养基中获得大量异源蛋白时常常出现问题。在此，我们提出一种使用夹心菌落来评估转基因菌株异源蛋白分泌情况的快速方法。通过在黑曲霉中表达双孢蘑菇的ABH1疏水蛋白，我们发现异源蛋白产量低可能是由于该蛋白在细胞外被蛋白水解降解。