实验动物模型

mTn-4xHA/lacZ TRTn3 terminal inverted repeatsHAHemagglutinin (HA) epitope loxRlox site target for Cre recombinaselacZ5'-truncated lacZ gene encoding beta-galactosidaseURA3URA3 gene from S. cerevisiaet ...

Mutagenizing a yeast gene with a mini-transposonFirst clone gene into vector pHSS6. Then:Day 1: Transform plasmid into bacteria expressing transposase. Day 2: Mate transformants to bacteria carrying m ...

Making library DNA from the DNA we send youThe library is distributed as individual pools in the form of DNA. You will be sent about a microgram of each pool available. Transform a suitable amount in ...

mTn-3xHA/lacZ TR Tn3 terminal inverted repeatsXaFactor Xa cleavage recognition siteloxRlox site target for Cre recombinaselacZ5'-truncated lacZ gene encoding beta-galactosidaseURA3URA3 gene from S. ce ...

mTn-lacZ/LEU2 TRTn3 terminal inverted repeatslacZ5'-truncated lacZ gene encoding beta-galactosidaseLEU2LEU2 gene from S. cerevisiaeampEncodes beta-lactamaseloxPlox site target for Cre recombinaseUses: ...

Quantitative Mating AssayGuthrie and Fink Guide to Yeast Genetics and Molecular Biology 19911. Grow a culture of the strain whose mating efficiency is to be determined to a density of about 1E7 cells/ ...

Replication timing by comparative hybridizationM. K. RaghuramanReference: Friedman K. L. M. K. Raghuraman W. L. Fangman and B. J. Brewer (1995) Analysis of the temporal program of replication initiati ...

Replication timing by density transfer M. K. Raghuraman 1. Grow cells at least 7 generations in0.1% (w/v) C-acetate or glucose 0.01% (w/v) N-ammonium sulfate Amino acids/supplements (as required added ...

UV Mutagenesis Materials:• Cells containing ADE3-URA3 plasmid grown in URA • dropout media • YEPD plates• sterile water• petri dishes• UV light source• “light-proof” boxProcedure:1) Grow cells 2-3 day ...

Replication timing using transient hemimethylationKatherine FriedmanReference: Friedman K. L. M. K. Raghuraman W. L. Fangman and B. J. Brewer (1995) Analysis of the temporal program of replication ini ...

Yeast Cell Cycle by Flow CytometrySusan Forsburg's method for Schizosaccharomyces pombe ReagentsCold absolute ethanol. 0.5 M Na citrate stock (filtered) 50mM diluted stock. 10 mg/ml RNase A (Boil 10 m ...

Actin Capture AssayDavid AmbergDialyze purified GST fusion proteins and actin into PBS + 1mM MgCl2 . Mix 5ug actin into 50ul total volume binding buffer. Mix 5ug GST-fusion protein into total volume 5 ...

Yeast IF without DehydrationDavid AmbergNote this protocol is a modified version of the protocol from Mark Rose in the CSH Yeast Genetics Course Manual.Grow cells at the appropriate temperature to 5x1 ...

问：如果诱饵蛋白对酵母细胞是有毒的，该怎么办？答：在有些情况下，在液体培养基中培养不好的菌珠可以在固体培养基上生长得很好。重悬克隆于1ml的SD/�Trp，接着将重悬液平铺于5 个100-mm的SD/�Trp平板。在30℃下温浴，直至平板上的克隆互相粘在一起。用5ml 0.5X YPDA刮下每块板上的克隆，并收集到一管中。接着就使用这个细胞重悬液进行正常的杂交反应。问：我的诱饵蛋白能直接激活报告基 ...

BacterioMatch 双杂交系统：在大肠杆菌中进行蛋白质相互作用分析简便快速。CytoTrap 双杂交系统：酵母细胞质中检测蛋白相互作用允许翻译后修饰和筛查转录激活因子/抑制因子HybriZAP 2.1酵母细胞核双杂交系统 ：采用l载体制备插入片段比率高，代表性好的cDNA/基因组文库，易于转化成质粒文库用于筛选同附件中是相关的参考资料。 点击浏览该文件 ...

　1 甲醇酵母表达系统的特点　　1.1 宿主　　七十年代巴斯德毕赤酵母曾被用于生产单细胞蛋白（SCP），有很好的发酵基础，菌体密度可达100g/L干重。其生长培养液的组分包括无机盐、微量元素、生物素、氮源和碳源，廉价而无毒。它能在以甲醇为唯一碳源的培养基中快速生长，其中醇氧化酶AOX――甲醇代谢途径的关键酶可达细胞可溶性蛋白的30％。而在葡萄糖、甘油或乙醇作为碳源的培养细胞中则检测不到AOX。AO ...

中文摘要：蛋白质分子间相互作用与识别是21世纪生命科学研究的前沿和热点. 分子对接方法是研究这一课题有效的计算机模拟手段. 通常，蛋白质-蛋白质分子对接包括四个阶段：搜索受体与配体分子间的结合模式，过滤对接结构以排除不合理的结合模式，优化结构，用精细的打分函数评价、排序对接模式并挑选近天然构象. 结合国内外研究蛋白质-蛋白质分子对接方法进展和本研究小组的工作，对以上四个环节做了详细的综 ...

1. Introduction and BackgroundThere is a great need for general methods to characterize the proteins that contemporary biology makes available. The list of such proteins needing further characterizati ...

ElectroporationDNA Preparation: 1. Cut the required amount of DNA (banded on a CsCl gradient) with the appropriate restriction enzyme and check for complete digestion by running 500 ng on a minigel. ...

酵母双杂交系统酵母双杂交系统是在真核模式生物酵母中进行的，研究活细胞内蛋白质相互作用，对蛋白质之间微弱的、瞬间的作用也能够通过报告基因的表达产物敏感地检测得到，它是一种具有很高灵敏度的研究蛋白质之间关系的技术。大量的研究文献表明，酵母双杂交技术既可以用来研究哺乳动物基因组编码的蛋白质之间的互作，也可以用来研究高等植物基因组编码的蛋白质之间的互作。因此，它在许多的研究领域中有着广泛的应用。 1、利用 ...