共查询到20条相似文献,搜索用时 15 毫秒
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丝状真菌的木质纤维素水解酶基因的表达很大程度上受到转录水平的调控,对纤维素酶系调控转录因子的研究对提高纤维素酶的产量具有重要意义。为了挖掘纤维素酶表达新调控基因,本研究对粗糙脉孢菌锌指转录因子C2H2家族的57株基因敲除突变株在以2%结晶纤维素为唯一碳源的培养条件下进行产纤维素酶水平分析筛选。通过测定发酵液的蛋白、内切β-1,4-葡聚糖酶酶活、外切纤维素酶酶活、β-葡萄糖苷酶酶活、木聚糖酶酶活及生物量,发现突变株L-38、L-85、L-40、L-64、L-99、L-07、L-86在蛋白水平及内切β-1,4-葡聚糖酶酶活水平均比野生型有25%-77%不等的显著提高,而突变株L-87、L-06在蛋白水平及内切β-1,4-葡聚糖酶酶活水平均比野生型有65%-80%不等的显著降低。这些纤维素酶新调控基因的获得为后续相关表达调控的研究提供了新素材。 相似文献
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Thedei G. Nozawa S.R. Simoes A.L. Rossi A. 《World journal of microbiology & biotechnology》1997,13(6):609-611
The mycelial Pi-repressible alkaline phosphatase of the wild-type strain 74A of Neurospora crassa was separated into at least ten isoforms by isoelectric focusing. The components visualized by activity with sodium -naphthyl phosphate as the substrate were predominantly acidic proteins with isoelectric points ranging from pH 4.5 to 7.6. The number of these isoforms was a function of growth pH. Strain pho-2A did not produce active Pi-repressible alkaline phosphatase (the pho-2 gene codes for its amino acid sequence), which gives an indication that these isoforms are encoded by the same structural gene. 相似文献
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L B Crotti H F Terenzi J A Jorge M L T M Polizeli 《Journal of industrial microbiology & biotechnology》1998,20(3-4):238-243
Pectolytic enzymes from the hyperproducer exo-1 mutant of Neurospora crassa are induced either by pectin or galactose. Galactose-induced pectinases, in contrast with pectin-induced enzymes, are not
affected by glucose repression. Here, the pectolytic enzymes induced by galactose were purified and characterized. Extracellular
pectolytic activities were separated into two main fractions. Pool I contained lyases, and a polygalacturonase (PG) copurifying
as a complex of about 80 kDa (gel filtration). Pool II contained PG only. Under urea-SDS-PAGE the lyases and polygalacturonase
from pool I migrated with an apparent MW of 56.2 kDa, and 34.3 kDa, respectively. PG from pool II exhibited an apparent MW
of 44.7 kDa. Cell extracts contained PG free of lyase activities. Purified intracellular PG migrated (SDS-PAGE) as a single
band of apparent MW of 31.5 kDa. All pectinases were glycoproteins (18.5–39% carbohydrate), with stability and optimum pH
at 5–6 and 9–10 for PG and lyases, respectively. Temperature optima were 40–50°C, respectively. All enzymes were inactivated
at 60°C, with a half-life from 1.5 to 5 min. Activation energy (Ea) values for extracellular and intracellular PG varied between
0.45 and 2.0 Kcal mol−1. Pool II and intracellular PG and lyases, exhibited a random mechanism of hydrolysis. Pool I PG exhibited an exo character.
Received 20 October 1997/ Accepted in revised form 28 February 1998 相似文献
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木质纤维素降解真菌粗糙脉孢菌天然具有吸收利用多种单糖和寡糖的能力,但是目前基因组中注释的预测糖转运蛋白仍然有过半功能未知。本研究从全基因组水平系统分析了粗糙脉孢菌预测糖转运蛋白的转运底物。研究发现两个转运蛋白(NCU01868和NCU08152)具有转运多种己糖底物的功能,因此分别命名为NcHXT-1和NcHXT-2。利用荧光共振能量转移技术(FRET)确认了NcHXT-1/-2具有葡萄糖转运功能。在己糖转运蛋白全缺酿酒酵母EBY.VW4000中分别过表达NcHXT-1/-2,能恢复其在葡萄糖、半乳糖或甘露糖的液体培养基中生长并生成乙醇的能力。NcHXT-1/-2在很多纤维素降解真菌中均具有保守的同源蛋白。本研究通过全基因组扫描鉴定,发现了两个保守的丝状真菌己糖转运蛋白,为真菌降解利用木质纤维素及酵母利用单糖发酵提供了新的改造靶点。 相似文献
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The nit-2 gene of Neurospora crassa encodes the major nitrogen regulatory protein which acts in a positive fashion to activate the expression of many different structural genes during conditions of nitrogen limitation. An E. coli-expressed NIT2/-Gal fusion protein binds specifically to DNA in vitro by recognizing GATA core elements. Nuclear extracts prepared from a wild-type N. crassa strain contain a protein factor which displays all of the properties expected for the native NIT2 protein. The native NIT2 protein in nuclear extracts binds with high affinity to DNA fragments which contain two GATA elements, weakly to fragments with a single GATA element, and fails to bind to DNAs which lack these sequences. The DNA binding ability of the protein factor in nuclear extracts is efficiently blocked by a polyclonal antibody developed against the zinc-finger region of NIT2 protein. Western blot analysis with the anti-NIT2 antiserum revealed a specific protein with a size of approximately 110,000 daltons, in excellent agreement with the predicted size of NIT2. Both the specific NIT2 DNA binding activity and the protein detected by Western blot are totally lacking in nuclear extracts of a nit-2 rip mutant strain. These results all support the conclusion that the native NIT2 protein in Neurospora cells has been identified. The NIT2 protein is localised in nuclei and could not be detected in the cytoplasmic fraction of cells subjected to nitrogen derepression or nitrogen repression, indicating that the nuclear import of NIT2 is not regulated. 相似文献
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OsSPX1 suppresses the function of OsPHR2 in the regulation of expression of OsPT2 and phosphate homeostasis in shoots of rice 总被引:2,自引:0,他引:2
Fang Liu Zhiye Wang Hongyan Ren Chenjia Shen Ye Li Hong‐Qing Ling Changyin Wu Xingming Lian Ping Wu 《The Plant journal : for cell and molecular biology》2010,62(3):508-517
Phosphate (Pi) homeostasis in plants is required for plant growth and development, and is achieved by the coordination of Pi acquisition, translocation from roots to shoots, and remobilization within plants. Previous reports have demonstrated that over‐expression of OsPHR2 (the homolog of AtPHR1) and knockdown of OsSPX1 result in accumulation of excessive shoot Pi in rice. Here we report that OsPHR2 positively regulates the low‐affinity Pi transporter gene OsPT2 by physical interaction and upstream regulation of OsPHO2 in roots. OsPT2 is responsible for most of the OsPHR2‐mediated accumulation of excess shoot Pi. OsSPX1 suppresses the regulation on expression of OsPT2 by OsPHR2 and the accumulation of excess shoot Pi, but it does not suppress induction of OsPT2 or the accumulation of excessive shoot Pi in the Ospho2 mutant. Our data also show that OsSPX1 is a negative regulator of OsPHR2 and is involved in the feedback of Pi‐signaling network in roots that is defined by OsPHR2 and OsPHO2. This finding provides new insight into the regulatory mechanism of Pi uptake, translocation, allocation and homeostasis in plants. 相似文献
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Ivone Un San Leong Andrew N. Shelling 《Biochemical and biophysical research communications》2010,396(4):817-824
Long QT syndrome is a disorder that is characterised by a prolonged QT-interval and can lead to fatal cardiac arrhythmias. Many animal models have been created to study congenital long QT syndrome. Of these, zebrafish models have involved targeting two different KCNH2 gene (long QT syndrome 2) orthologues, termed zerg-2 and zerg-3, with differing cardiac phenotypes. In order to clarify this situation, this study uses a bioinformatic approach to search the current zebrafish genome sequence (Zv7 and Zv8 builds) to investigate and locate all likely zebrafish orthologues of the human KCNH2 gene. Quantitative real-time RT-PCR was also used to determine the temporal and spatial gene expression profile of the zebrafish orthologues. The data support the conclusion that zerg-2 and zerg-3 are apparent orthologues of different human genes encoding potassium ion channels, but that their functions have switched compared to the respective human proteins. 相似文献
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Cells of Saccharomyces cerevisiae grown in media with an initial pH of 2.5–6.0, acidified with a strong acid (HCl), exhibited the highest plasma membrane H+-ATPase-specific activity at an initial pH of 6.0. At a lower pH (above pH 2.5) ATPase activity (62–83% of the maximum level)
still allowed optimal growth. At pH 2.5, ATPase activity was about 30% of the maximum value and growth was impaired. Quantitative
immunoassays showed that the content of ATPase protein in the plasma membrane was similar across the entire pH range tested,
although slightly lower at pH 2.5. The decrease of plasma membrane ATPase activity in cells grown at low pH was partially
accounted for by its in vitro stability, which decreased sharply at pH below 5.5, although the reduction of activity was
far below the values expected from in vitro measurements. Yeast growth under acid stress changed the pattern of gene expression
observed at optimal pH. The level of mRNA from the essential plasma-membrane-ATPase-encoding gene PMA1 was reduced by 50% in cells grown at pH 2.5 as compared with cells grown at the optimal pH 5.0, although the content of ATPase
in the plasma membrane was only modestly reduced. As observed in response to other kinds of stress, the PMA2 promoter at the optimal pH was up to eightfold more efficient in cells grown at pH 2.5, although it remained several hundred
times less efficient than that of the PMA1 gene.
Received: 22 April 1996 / Accepted: 6 August 1996 相似文献
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Yeying Sun a b Quanqi Zhang a Jie Qi a Yanjie Chen a Qiwang Zhong a Chunmei Li a Yan Yu a Shuo Li a Zhigang Wang a a Key Laboratory of Marine Genetics Breeding Ministry of Education Qingdao China b College of Pharmacy Binzhou Medical University Yantai China 《Acta Genetica Sinica》2010,(2)