谢志雄
微生物遗传学
个性化签名
- 姓名:谢志雄
- 目前身份:
- 担任导师情况:
- 学位:
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学术头衔:
博士生导师
- 职称:-
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学科领域:
遗传学
- 研究兴趣:微生物遗传学
谢志雄,博士,教授
研究方向:微生物遗传学
学习与工作经历:
1987.9-1991.6 武汉大学生物系 生物化学专业 学士;
1991.7-1993.8 武汉市东西湖区农业科学技术研究所 助研;
1993.9-1996.6 武汉大学生命科学学院 动物学专业 硕士;
1999-1999.6 武汉大学生命科学学院 遗传学专业 博士;
1999.7至今 武汉大学生命科学学院教师
2002年晋升为副教授;2007年晋升为教授,2008年遴选为博士生导师;
1999.12-2002.8 武汉大学化学与分子科学学院 化学博士后流动站 博士后;
2003.3-2004.3 韩国 全北大学校 基础科学研究所 Research Associate。
近五年来承担的科研课题
国家自然科学基金项目CdSe/ZnS量子点的细胞生物效应研究(20677044)2007-2009主持28万元
国家自然科学基金项目细菌释放DNA与生物膜形成相关性研究(30370017)2004-2006主持20万元
国家自然科学基金项目环境中DNA的动态分析方法研究(20207005)2003-2005主持21万元
国家高技术研究发展计划(863计划)量子点等纳米材料环境污染检测技术基础(2007AA06Z407)2008-2010副组长25.38万元
国家自然科学基金创新研究群体新型生物医学探针技术基础及应用(20621502)2007-2009骨干20万元
近五年来获得的学术奖励
2007年“DNA表面化学及基于DNA的生物传感”获教育部自然科学奖一等奖。(排名第6)
2006年“Yeast transformation process studied by fluorescence
labeling technique”获湖北省自然科学优秀学术论文二等奖。(排名第1)
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成果阅读
480
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成果数
7
【期刊论文】Living Yeast Cells as a Controllable Biosynthesizer for Fluorescent Quantum Dots
谢志雄
Adv. Funct. Mater. 2009, 19, 2359-2364,-0001,():
-1年11月30日
There are currently some problems in the field of chemical synthesis, such as environmental impact, energy loss, and safety, that need to be tackled urgently. An interdisciplinary approach, based on different backgrounds, may succeed in solving these problems. Organisms can be chosen as potential platforms for materials fabrication, since biosystems are natural and highly efficient. Here, an example of how to solve some of these chemical problems through biology, namely, through a novel biological strategy of coupling intracellular irrelated biochemical reactions for controllable synthesis of multicolor CdSe quantum dots (QDs) using living yeast cells as a biosynthesizer, is demonstrated. The unique fluorescence properties of CdSe QDs can be utilized to directly and visually judge the biosynthesis phase to fully demonstrate this strategy. By such a method, CdSe QDs, emitting at a variety of single fluorescence wavelengths, can be intracellularly, controllably synthesized at just 30-C instead of at 300-C with combustible, explosive, and toxic organic reagents. This green biosynthetic route is a novel strategy of coupling, with biochemical reactions taking place irrelatedly, both in time and space. It involves a remarkable decrease in reaction temperature, fromaround 300℃ to 30℃and excellent color controllability of CdSe photoluminescence. It is well known that to control the size of nanocrystals is a mojor challenge in the biosynthesis of high-quality nanomaterials. The present work demonstrates clearly that biological systems can be creatively utilized to realize controllable unnatural biosynthesis that normally does not exist, offering new insights for sustainable chemistry.
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【期刊论文】Quantum Dot-Ferrichrome Bioprobes for Recognition of Pseudomonas fluorescens
谢志雄
J. Phys. Chem. C 2009, 113, 9169-9174,-0001,():
-1年11月30日
Ferrichrome is a kind of high-affinity iron(III) chelators and its complex with Fe3+ can be specifically recognized by the corresponding receptors located on the outer membrane of some bacterial cells. Quantum dots as a type of semiconductor nanocrystals have high luminance and good resistance to photobleaching. Herein, ferrichrome was conjugated with polyethylene glycol-phosphoethanolamine-coated quantum dots to produce a new type of quantum dot-ferrichrome bioprobes. The quantum dot-ferrichrome bioprobes were used for recognizing Pseudomonas fluorescens (P. fluorescens) isolated from Dong-Hu Lake in China. P. fluorescens could be recognized quickly and sensitively with quantum dot-ferrichrome bioprobes by forming cell clusters through reaction between ferrichrome-ferric complex and its receptors. It was multiple ligands on a quantum dot and multiple receptors on a P. fluorescens cell that made the bacteria cluster. Single cells can also be realized with superfluous quantum dot-ferrichrome bioprobes to occupy the receptors on bacterial outer membrane. The biosensitive quantum dot-ferrichrome kept excellent fluorescent property of quantum dot and might be a promising bioprobes for fluorescent Pseudomonads targeting.
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谢志雄
JOURNAL OF BACTERIOLOGY, Feb. 2009, p. 713-719,-0001,():
-1年11月30日
Spontaneous plasmid transformation of Escherichia coli occurs on nutrient-containing agar plates. E. coli has also been reported to use double-stranded DNA (dsDNA) as a carbon source. The mechanism(s) of entry of exogenous dsDNA that allows plasmid establishment or the use of DNA as a nutrient remain(s) unknown. To further characterize plasmid transformation, we first documented the stimulation of transformation by agar and agarose. We provide evidence that stimulation is not due to agar contributing a supplement of Ca2, Fe2, Mg2, Mn2, or Zn2. Second, we undertook to inactivate the E. coli orthologues of Haemophilus influenzae components of the transformation machine that allows the uptake of single-stranded DNA (ssDNA) from exogenous dsDNA. The putative outer membrane channel protein (HofQ), transformation pseudopilus component (PpdD), and transmembrane pore (YcaI) are not required for plasmid transformation. We conclude that plasmid DNA does not enter E. coli cells as ssDNA. The finding that purified plasmid monomers transform E. coli with single-hit kinetics supports this conclusion; it establishes that a unique monomer molecule is sufficient to give rise to a transformant, which is not consistent with the reconstitution of an intact replicon through annealing of partially overlapping complementary ssDNA, taken up from two independent monomers. We therefore propose that plasmid transformation involves internalization of intact dsDNA molecules. Our data together, with previous reports that HofQ is required for the use of dsDNA as a carbon source, suggest the existence of two routes for DNA entry, at least across the outer membrane of E. coli.
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【期刊论文】Visualized investigation of yeast transformation induced with Li+and polyethylene glycol
谢志雄
Talanta 77 (2008) 262-268,-0001,():
-1年11月30日
The effects of Li+ and polyethylene glycol (PEG) on the genetic transformation of Saccharomyces cerevisiae were investigated by using fluorescence microscopy (FM) to visualize the binding of plasmid DNA labeled with YOYO-1 to the surface of yeast cells, scanning electron microscopy (SEM) and atomic force microscopy (AFM) to image the change in surface topography of yeast cells, coupled with transformation frequency experiments. The results showed that under the same conditions, the transformation frequencies of yeast protoplastswere much higher than those of intact yeast cells. PEGwas absolutely required for the binding of DNA to the surface of intact yeast cells or yeast protoplasts, and had no effect on the surface topography of intact yeast cells or yeast protoplasts. In the presence of PEG, Li+ could greatly enhance the binding of plasmid DNA to the surface of intact yeast cells, increase their transformation frequency, and affect their surface topography. On the other hand, no effect on the DNA binding to the surface of protoplasts and no increase in the number of transformants and no surface topography changes were found upon the treatment with Li+ to protoplasts. In the present work, the effects of Li+ and PEG on yeast genetic transformation were directly visualized, rather than those deduced from the results of transformation frequencies. These results indicate that cell wall might be a barrier for the uptake of plasmid DNA. Li+could increase the permeability of yeast cell wall, then increase the exposed sites of DNA binding on intact yeast cells. The main role of PEG was to induce DNA binding to cell surface
Transformation, Li+, , PEG, Yeast, Protoplast, AFM
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【期刊论文】Escherichia coli is naturally transformable ina novel transformation system
谢志雄
FEMS Microbiol Lett 265 (2006) 249-255,-0001,():
-1年11月30日
A novel transformation system, in which neither a nonphysiological concentration of Ca2+and temperature shifts nor electronic shocks were required, was developed to determine whether Escherichia coli is naturally transformable. In the new protocol, E. coli was cultured normally to the stationary phase and then cultured statically at 37℃ in Luria-Bertani broth. After static culture, transformation occurred in bacteria spread on Luria-Bertani plates. The protein synthesis inhibitor chloramphenicol inhibited this transformation process. The need for protein synthesis in plated bacteria suggests that the transformation of E. coli in this new system is regulated physiologically.
Escherichia coli, natural transformation, static culture
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【期刊论文】高产铁载体荧光假单胞菌Pseudomonas fluorescens sp2f的筛选鉴定及其铁载体特性研究
谢志雄, ZHAO Xiang, CHEN Shaoxing, XIE Zhixiong, SHEN Ping
微生物学报:2006,46 (5):691~695,-0001,():
-1年11月30日
采用改进的CAS 检测平板从东湖中筛选得到了一株高产铁载体细菌sp2f,并用CAS检测液定量检测其分泌铁载体量,发现其AsPAr仅0109(OD680),Su(Siderophore Unit)为90%,达到产铁载体菌最高级。用BIOLOG检测板,结合细菌生理生化反应、形态观察和16S rDNA 序列比对分析等分类鉴定方法,确定sp2f为一株荧光假单胞菌。P. fluorescens sp2f生长过程中胞外铁载体的量在对数生长前期累积达到最高后有所减少,至稳定期时菌液中铁载体量达到稳定。在已知铁载体特异吸收峰波长下,用反向高效液相色谱检测无铁环境和高铁环境下培养液上清,比较发现sp2f 上清含有3种含儿茶酚胺类基团铁载体,其中包括荧光和非荧光性的脓菌素,200μmolPL Fe2+可完全抑制荧光性质脓菌素的分泌,但非荧光脓菌素的分泌不受抑制,并且对非脓菌素的儿茶酚胺类铁载体的合成分泌反而具有一定的诱导作用。
高产铁载体, 荧光假单胞菌, 细菌鉴定, 脓菌素
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【期刊论文】Yeast Transformation Process Studied by Fluorescence Labeling Technique
谢志雄
Bioconjugate Chem. 2005, 16, 250-254,-0001,():
-1年11月30日
A new method based on fluorescence imaging and flow cytometry was developed to investigate the transformation process of Saccharomyces cerevisiae AY. Yeast and fluorescent-labeled plasmid pUC18 were used as models of cells and DNA molecules, respectively. Binding of DNA molecules to yeast cell surfaces was observed. Factors influencing DNA binding to cell surfaces were investigated. It has been found that poly(ethylene glycol) (PEG) could induce DNA binding to yeast surfaces, while Li+ showed a weak effect on the binding. When both Li+ and PEG were used, synergetic effect occurred, resulting in the binding of pUC18 to the surface of more yeast cells compared with that in the presence of PEG or Li+ only. It was also confirmed that heat shock, Li+, and PEG all can increase the permeability of yeast cells. This simple method is helpful for understanding the process of yeast transformation and can be used to investigate the interaction of DNA with cell surfaces.
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