汪天虹
从事微生物遗传与分子生物学研究,侧重于细菌及丝状真菌纤维素、半纤维素降解、代谢酶的酶学及分子生物学研究。
个性化签名
- 姓名:汪天虹
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学术头衔:
博士生导师
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学科领域:
微生物学
- 研究兴趣:从事微生物遗传与分子生物学研究,侧重于细菌及丝状真菌纤维素、半纤维素降解、代谢酶的酶学及分子生物学研究。
汪天虹,山东大学微生物技术国家重点实验室教授,博士生导师,分子技术研究室主任。申请者所领导的研究室长期从事微生物遗传与分子生物学研究,侧重于细菌及丝状真菌纤维素、半纤维素降解、代谢酶的酶学及分子生物学研究。在丝状真菌酶学、遗传学和分子生物学研究方面有多年的工作基础,曾先后到澳大利亚悉尼大学农化系和芬兰国家生物技术研究所(VTT)深造和进行微生物分子生物学研究工作。近年来承担和参加了18项相关内容的国家和省部级项目,在国内外重要学术刊物上刊出研究论文50余篇(SCI论文20篇)。对瑞氏木霉纤维素、半纤维素降解代谢酶基因克隆与表达、酶结构域研究、蛋白质随机诱变和定向进化、丝状真菌原生质体制备与转化、强表达整合型载体的构建、表达系统宿主菌的改造、基因一步置换与定位整合,通过基因寻靶技术构建定位基因突变体、纤维素、半纤维素酶启动子的人工改造、转录因子研究、基因的反义抑制、代谢途径改造等多方面都有成功的经验和突出的研究成果。几个主要研究方向包括:丝状真菌遗传与分子生物学研究;海洋微生物适冷酶酶学与分子生物学;生防木霉的分离与遗传改造等。参加“微生物对木质纤维素降解机理的研究”,获国家教育部基础类研究1等奖(2001年);“纤维素、半纤维素降解代谢酶的分子生物学研究” 获山东省科委科技进步3等奖(2001年,首位)。主编”微生物分子育种原理与技术” (化工出版社 2005.8月 北京) 参编”环境生物工程” (化学工业出版社 2002.9月 北京)
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515
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成果数
10
汪天虹, Tian-Hong WANG*, Ti LIU, Zhi-Hong WU, Shi-Li LIU, Yi LU, and Yin-Bo QU
Acta Biochimica et Biophysica Sinica 2004, 36 (10): 667-672,-0001,():
-1年11月30日
To construct strains of the filamentous fungus Trichoderma reesei with low cellobiohydrolases while high endoglucanase activity, the Pcbh1-eg3-Tcbh1 cassette was constructed and the coding sequence of the cellobiohydrolase I (CBHI) gene was replaced with the coding sequence of the eg3 gene by homologous recombination. Disruption of the cbh1 gene was confirmed by PCR, Southern dot blot and Western hybridization analysis in two transformants denoted as L13 and L29. The filter paper-hydrolyzing activity of strain L29 was 60% of the parent strain Rut C30, and the CMCase activity was increased by 33%. This relatively modest increase suggested that the eg3 cDNA under the control of the cbh1 promoter was not efficiently transcribed as the wild type cbhl gene. However our results confirmed that homologous recombination could be used to construct strains of the filamentous fungus Trichoderma reesei with novel cellulase profile. Such strains are of interest from the basic science perspective and also have potential industrial applications.
Trichoderma reesei, gene replacement, gene disruption, cbh1, eg3
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汪天虹, Wei Huang, Tian-Hong Wang, ∗, JingWu & Chun-Qiang Liu-
Biotechnology Letters 24: 395-399, 2002.,-0001,():
-1年11月30日
A mitochondrial gene cluster, encoding proteins homologous to NADH dehydrogenase subunits II and III (ND2 and ND3) and seven tRNAs, from Trichoderma reesei QM9414 was cloned and sequenced. These genes are clustered tandemly on the mitochondrial genome of QM9414. Phylogenetic analysis showed that ND2 and ND3 were most closely related to the mitochondrialND subunits II (71% identity) and III (70% identity) from Podospora anserine. Northern dot blot analysis showed that the nd-and nd3 genes are actively transcribed in the T. reesei mitochondria.
mitochondria, NADH dehydrogenase subunits, sequence analysis, Trichoderma reesei, tRNA
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汪天虹, WAN G Tian-hong, ZHAN G Gang and HOU Yun-hua
CHEM. RES. CHINESE U.2004, 20 (1), 60-64,-0001,():
-1年11月30日
The filamentous fungi from the Huanghai sea sludge were screened according to their ability to produce cold-ac-tive α-amylase. The strain with the highest amylase activity was identified as Penicillium species. The α-amylase purified by ammonium sulphate precipitation and column chromatography on DEAE2sepharose and sephadex G2100 shows a molecular weight of about 55000 and a p I of 4. 38. The enzyme is stable in a pH range of 5. 5-8. 0 and has a maximum activity at pH 6. 0. Compared with the α-amylase from mesophiles and thermophiles, the cold-ac-tive enzyme shows a high enzyme activity at lower temperatures and a high sensitivity at temperatures higher than 50℃. The optimal temperature is 40℃and the activity decreases dramatically at temperatures above 50℃. Ca2+shows a significant effect on maintaining the structure and the activity of the enzyme. EDTA and Cu2 + are its in-hibitors. The products from the hydrolysis of soluble starch with the cold-active enzyme are maltose and other oligosaccharides.
Cold2activeα-amylase, Marine Penicillium, Purification, Characterization
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汪天虹, J. Liu, S.-Y. Sun and T.-H. Wang
Letters in Applied Microbiology 2004, 38, 277-282,-0001,():
-1年11月30日
To construct a yeast one-hybrid system and isolate transcriptional activators. Methods and Results: A 1
ACE II, promoter, trans, c, r, i, p, t, ion activator, T., reesei,, yeast one-hybrid-system,, xyn2.,
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【期刊论文】Antisense inhibition of xylitol dehydrogenase gene, xdh1 from Trichoderma reesei
汪天虹, T.H. Wang, Y.H. Zhong, W. Huang, T. Liu and Y.W. You
Letters in Applied Microbiology 2005,-0001,():
-1年11月30日
To inhibit xylitol dehydrogenase (XDH) in Trichoderma reesei by antisense inhibition strategy and construct novel strains capable of accumulating xylitol. Methods and Results: The xdh1 antisense expression plasmid pGTA-xdh was constructed by inserting xdh1 DNA fragment inversely between the gpdA promoter and the trpC terminator from Aspergillus nidulans into a pUC19 plasmid backbone. Trichoderma reesei protoplasts were co-transformated with pGTA-xdh and hygromycin B resistance plasmid pAN7-1. Of 20 transformants screened from the selective medium, one transformant with the highest xylitol accumulation, designated ZY15, showed a distinct reduction (c. 52%) in XDH activity compared with the original strain Rut-C30. The results of Southern hybridization and PCR assay showed that the antisense expression cassette of xdh1 was integrated into the genome of T. reesei. The RT-PCR analysis proved that antisense RNA effectively inhibited XDH expression (c. 65%). Xylitol accumulation 2
antisense inhibition, Trichoderma reesei, xdh1,, xylitol, xylitol dehydrogenase.,
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汪天虹, Yun-Hua HOU, Tian-Hong WANG*, Hao LONG, and Hui-Yuan ZHU
Acta Biochimica et Biophysica Sinica 2006, 38 (2): 1672-9145,-0001,():
-1年11月30日
A novel cold-adaptive xylanolytic Penicillium strain FS010 was isolated from Yellow sea sediments. The marine fungus grew well from 4 to 20
marine fungi, Penicillium chrysogenum, cold-active xylanase, gene expression
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汪天虹, Yao Hua Zhong & Xiao Li Wang & Tian Hong Wang & Qiao Jiang
Appl Microbiol Biotechnol DOI 10.100,-0001,():
-1年11月30日
Filamentous fungus Trichoderma reesei QM9414 was successfully transformed with Agrobacterium tumefaciens AGL-1 for random integration of transforming DNA (T-DNA). Co-cultivation of T. reesei conidia or protoplasts with A. tumefaciens in the presence of acetosyringone resulted in the formation of hygromycin B-resistant fungal colonies with high transformation frequency. Nine randomly selected resistant clones were proved to be stable through mitotic cell division. The integration of the hph gene into T. reesei genome was determined by PCR and dot blot analysis. Transgenic T. reesei strains were analyzed using TAIL-PCR for their T-DNA contents. The results showed that T-DNA inserts occurred evidently by fusing DNA at T-DNA borders via random recombination, which suggests that Agrobacterium-mediated transformation is a potentially powerful tool towards tagged mutagenesis and gene transfer technology for T. reesei.
Agrobacterium-mediated transformation, Trichoderma reesei, T-DNA., Insertional mutagenesis, hygromycin B resistance, TAIL-PCR
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汪天虹, Merja Penttil, 高培基, 王春卉, 钟玲
生物工程学报,1998,14(3):320~325,-0001,():
-1年11月30日
将在木聚糖上生长的瑞氏木霉(Trichodermareesei)RutC230的cDNA文库全部质粒转化已携带有毕赤氏酵母(Pichiastipitis)木糖还原酶基因的重组酿酒酵母(Saccharomycescerevisiae)菌株H475,在H475中构建了瑞氏木霉的CDNA表达亚文库。在以木糖为唯一碳源的选择性酵母合成培养基上,从该亚文库中筛选到瑞氏木霉木糖醇脱氢酶CDNA基因,该基因片段长为113kbSouthern、Northern印迹杂交分析和蛋白质凝胶电泳结果表明该基因确实来源于瑞氏木霉,所编码蛋白质分子量约为40kDa,携带有毕赤氏酵母木糖还原酶和瑞氏木霉木糖醇脱氢酶基因的重组酵母能够在以木糖为唯一碳源的培养基上生长,并能将90%以上的木糖转化为木糖醇、乙醇和其它副产品。
瑞氏木霉,, 木糖醇脱氢酶基因,, 酿酒酵母,, 木糖
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【期刊论文】瑞氏木霉木糖代谢关键酶基因在不同碳源条件下的表达*
汪天虹, Merja Penttil, 高培基
微生物学报,1999,36(12):503~509,-0001,():
-1年11月30日
用20种不同的碳源(包括单一碳源和混合碳源)分别培养瑞氏木霉(Trichodermareesei)QM9414。通过一系列Northern杂交分析检测瑞氏木霉木糖还原酶(XR),木糖醇脱氢酶(XDH)以及转醛醇酶(TAL)mRNA的表达情况。实验结果证实,槐糖和木二糖是xr和xdh表达的强诱导物,阿拉伯糖和乳糖也有较强的诱导作用。葡萄糖在培养基中的存在阻遏该二基因的表达。当葡萄糖耗尽以后,培养基中不存在任何诱导物的情况下,xr和xdh以一定的基础水平进行转录。相比较,tal基因在每种碳源上都是强表达。
瑞氏木霉, 木糖还原酶基因, 木糖醇脱氢酶基因, Northern 杂交, 基因表达
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汪天虹, 吴志红, 刘世利, 曲音波
中国生物化学与分子生物学报,2003,19(6):736~745,-0001,():
-1年11月30日
采用PCR技术体外扩增获得了瑞氏木霉外切葡聚糖纤维二糖水解酶(CBH)启动子和终止子序列。并以大肠杆菌质粒PUC19为骨架,在该启动子和终止子序列间加入多克隆位点,构建了瑞氏木霉强表达整合型载体PTRIL。以质粒PAN721为模板,体外扩增了带有潮霉素磷酸转移酶(hph)基因的DNA片段,将hph插入PTRIL的cbh1启动子和终止子序列之间,构建了Pcbh1-hph-Tcbh1表达盒。用此表达盒转化瑞氏木霉C30原生质体,在潮霉素平板上得到15株抗性转化子。对其中的H1转化子进行了PCR和Southern印迹分析,证实hph基因确实整合到转化子染色体DNA上,并在Pcbh1启动子控制下进行高效表达。转化子H1对潮霉素抗性达150mgPL,比出发菌株提高2倍.瑞氏木霉强表达整合型载体PTRIL的构建成功为开展瑞氏木霉分子生物学研究以及进一步的工程菌株构建工作奠定了基础。
瑞氏木霉,, 外源基因表达系统,, cbh1启动子,, 构建与表达
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