h**********r
发帖数: 671 | 1 Title:
Gene Disruption in the Budding Yeast Saccharomyces cerevisiae
Series: Methods in Molecular Biology | Volume: 313 |
my email is m******[email protected], Thank you so much! |
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t****a
发帖数: 439 | 2 Key words:
Ethanol
Elimination of glycerol production,
Redox balance
Stress management
Saccharomyces cerevisiae
Please drop me a message ONLY if you are an expert in Saccharomyces
cerevisiae metabolic engineering.
Thanks. |
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y******y
发帖数: 107 | 3 yeast有两大类， 一是Saccharomyces cerevisiae，二是 Schizosaccharomyces pombe。
Saccharomyces cerevisiae 主要有三种，1, mating type a haploid； 2, mating
type alpha haploid； 3, diploid (这个是一种还是两种？）。
那么Schizosaccharomyces pombe 有几种类型呢？ google了，但是没有发现有价值的
结果。谢谢 |
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h**********r
发帖数: 671 | 4 我在实验室用的Thermo，貌似work的不是很好。
Yeast DNA Extraction
(The protocol was modified by housepainter on December 28, 2009 according to
manufacture’s instructions)
Yeast DNA Extraction Kit, Thermo Number Description: 78870
Storage: Store at 4°C; if precipitant has formed, gently warm DNA Releasing
Reagents A and B at 37°C for 1-5 minutes.
Kit Contents:
Y-PER Reagent, 25 ml
DNA Releasing Reagent A, 20 ml
DNA Releasing Reagent B, 20 ml
Protein Removal Reagent, 10 ml
1. Pellet a 10 ml S. cerevisiae culture gr... 阅读全帖 |
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a****o
发帖数: 1786 | 5 I assume it is not S. cerevisiae, if it is Yarrowia, you can design PCR
primers for matA1, matA2 and matB1, matB2 and perform colony PCR. If it only
gives product for A or B, then it is haploid; if it gives product for both
A and B, it is diploid.
If it is S. cerevisiae, then you should not have called it non-conventional
yeast. Then you can plate lawns of tester strains and streak/replicate you
strain on them.If you strain inhibits growth of either tester strain, it is
haploid with the opposite... 阅读全帖 |
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A********e
发帖数: 354 | 6 【 以下文字转载自 Biology 讨论区 】
发信人: albertsmwk (.)(.), 信区: Biology
标 题: 第一批“青年千人计划”生物类@publication列表@
发信站: BBS 未名空间站 (Thu Aug 18 15:04:16 2011, 美东)
花了一个小时，深深的鄙视一下自己的无聊行径！！
125 蔡亮 男 1980年11月 复旦大学 生命
科学 2007年12月毕业于[美国]北卡大学 [美国]加州大学旧金山分校 博士后
Cai L, Mostov K. Polarity is destiny. Cell. 2009 Nov 13;139(4):660-2. PubMed
PMID: 19914162; PubMed Central PMCID: PMC2900917.
Cai L, Makhov AM, Schafer DA, Bear JE. Coronin 1B antagonizes cortactin and
remodels Arp2/3-containing actin branche... 阅读全帖 |
|
c**i
发帖数: 6973 | 7 (1) Benjamin Yeh, From hunger to Honour: Top Taiwan Baker Thanks Mum. AFP,
Sept 8, 2011
http://news.yahoo.com/hunger-honour-top-
taiwan-baker-thanks-mum-042647138.html
("Japan ran Taiwan as a colony for 50 years after 1895 and introduced the
island to bread, albeit in a soft version without hard crusts")
Note:
(a) WU Pao-chun 吳 寶春
Loa Iok-sin, Taiwanese Baker Wins Industry’s Top Prize in Paris. Taipei
Times, Mar 14, 2011.
http://www.taipeitimes.com/News
/front/archives/2010/03/14/2003467997
(b)... 阅读全帖 |
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w***g
发帖数: 5958 | 8 我家一直用sams club/cost卖的那种超大包的instant dry yeast，感觉用一辈子都够
了的那种。发面的时候一般都是用面包机。自己确实不太好控制温度。糖和盐不是必须
的，直接加水和面混在一起就可以。
做面包/酿啤酒/酿葡萄酒的都是一种叫Saccharomyces cerevisiae的yeast中的最重要
的species的不同的strains，功能都差不多，可能口味有细微差别。我试过用发面包的
酵母酿酒，也能酿出来。 |
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p*******g
发帖数: 2976 | 9 【 以下文字转载自 Biology 讨论区 】
发信人: albertsmwk (.)(.), 信区: Biology
标 题: 第一批“青年千人计划”生物类@publication列表@
发信站: BBS 未名空间站 (Thu Aug 18 15:04:16 2011, 美东)
花了一个小时，深深的鄙视一下自己的无聊行径！！
125 蔡亮 男 1980年11月 复旦大学 生命
科学 2007年12月毕业于[美国]北卡大学 [美国]加州大学旧金山分校 博士后
Cai L, Mostov K. Polarity is destiny. Cell. 2009 Nov 13;139(4):660-2. PubMed
PMID: 19914162; PubMed Central PMCID: PMC2900917.
Cai L, Makhov AM, Schafer DA, Bear JE. Coronin 1B antagonizes cortactin and
remodels Arp2/3-containing actin branche... 阅读全帖 |
|
d*******u
发帖数: 5337 | 10 Hi reviewers,
I wish you all have a happy new year.
Please check the following manuscripts to Biomedicine and pharmacotherapy.
If you are interested in reviewing these manuscripts, send your background
and Ms #. Feel free to pass this information to your friends and collogues.
Best regards,
xxxx
Executive editor
Biomedicine and Pharmacotherapy
1. Antiproliferative activity of Corylus avellana L. extracts against
human cancer cell lines
2. Poloxamer Surface modified Trimethyl Chitosan Nano... 阅读全帖 |
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s*******3
发帖数: 14 | 11 曾经协助老板审稿8篇，包括Biotech for Biofuel, Metabolic Engineering, PNAS和
Nature子刊等。自己独立有Biotech相关的会议审稿经历。
熟悉的领域；Saccharomyces cerevisiae, metabolic engineering, synthetic
biology, RNAi, CRISPR, cellulosic biofuel, yeast surface display, genome
engineering, directed evolution, high throughput screening等
如需要更为详细信息，请PM或者邮件s*******[email protected]。非常感谢您的帮助：） |
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k****r
发帖数: 593 | 12 【 以下文字转载自 Pharmaceutical 讨论区 】
发信人: kenter (sphinx), 信区: Pharmaceutical
标 题: 版上有没有哪位药厂工作的可以推荐 Summer Intern for Process Development Engineer
发信站: BBS 未名空间站 (Thu Feb 3 14:44:50 2011, 美东)
本人估计还有一年毕业，化工专业，主要是抗体蛋白质提纯方向。
•Specialized in protein purification process including chromatography
(AC, IEC, SEC, HIC), filtration (TFF, UF, DF, NF), precipitation, etc.
•Specialized in protein quantification including ELISA, SDS-PAGE,
Western blot, densitometry analysis, BCA, Bradford, enzymatic a... 阅读全帖 |
|
O***n
发帖数: 13127 | 13 【 以下文字转载自 Biology 讨论区 】
发信人: albertsmwk (.)(.), 信区: Biology
标 题: 第一批“青年千人计划”生物类@publication列表@
发信站: BBS 未名空间站 (Thu Aug 18 15:04:16 2011, 美东)
花了一个小时，深深的鄙视一下自己的无聊行径！！
125 蔡亮 男 1980年11月 复旦大学 生命
科学 2007年12月毕业于[美国]北卡大学 [美国]加州大学旧金山分校 博士后
Cai L, Mostov K. Polarity is destiny. Cell. 2009 Nov 13;139(4):660-2. PubMed
PMID: 19914162; PubMed Central PMCID: PMC2900917.
Cai L, Makhov AM, Schafer DA, Bear JE. Coronin 1B antagonizes cortactin and
remodels Arp2/3-containing actin branche... 阅读全帖 |
|
d********a
发帖数: 1947 | 14 在做面包的aisle找sacchromyces cerevisiae的active dry yeast，这个和酒曲有点不
同，算ale yeast，不过也可以发酵做酒酿用，但是发酵时间要就一点，还要先proof一
下，我自己做还会再多加勺糖，不过我喜欢甜一点的，发酵温度差不多70F，过了98F酵
母会死掉 |
|
b*****h
发帖数: 61 | 15 1.Journal: Biotechnology and Bioengineering
Volume 36 Issue 11, Pages 1133 - 1140
Title: High-level recombinant protein production in bioreactors using the
baculovirus-insect cell expression system
Author: Antoine W. Caron, Jean Archambault, Bernard Massie *
2. Journal:Methods in Enzymology
Volume 424, 2007, Pages 319-331
Title: Large-Scale Overexpression and Purification of ADARs from
Saccharomyces cerevisiae for Biophysical and Biochemical Studies
Author: Mark R. Macbetha and Brenda L. Bassa
多 |
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D*****t
发帖数: 1303 | 16 好像木看到跟蛋有关的成分啊？blue buffalo的就是这个wilderness的好，其他的都不太好
http://bluebuffalo.com/products/cats/wilderness-cat-dry.shtml
Ingredients
Deboned Chicken, Chicken Meal, Turkey Meal, Potato Starch, Fish Meal(natural source of Omega 3 Fatty Acids), Chicken Fat (naturally preserved with Mixed Tocopherols), Peas, Whole Potatoes, Flaxseed (natural source of Omega 3 and 6 Fatty Acids), Natural Chicken Flavor, Alfalfa Meal, Whole Carrots, Whole Sweet Potatoes, Cranberries, Blueberries, Barley Grass, Dried Parsley,... 阅读全帖 |
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c*******t
发帖数: 168 | 17 我们家吃的是这个spa select，把成分找出来了，这款果然是有鸡蛋的。
为什么说只有wildness的好其他都不好阿，我看配料都差不多似得阿。
http://www.petco.com/product/102979/Spa-Select-Adult-Chicken-and-Brown-Rice-Recipe-Cat-
Food.aspx?CoreCat=CatFFB_BlueBuffalo
不太好
natural source of Omega 3 Fatty
Acids), Chicken Fat (naturally preserved with Mixed Tocopherols), Peas,
Whole Potatoes, Flaxseed (natural
source of Omega 3 and 6 Fatty Acids), Natural Chicken Flavor, Alfalfa Meal,
Whole Carrots, Whole Sweet
Potatoes, Cranberries, Blueberries, Barley Grass, Dried... 阅读全帖 |
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v***6
发帖数: 199 | 18 两小吃完babycat 34吃babycat36，吃了有2个多月，不爱喝水。后来一猫有阵子尿团巨
大，另一猫尿尿时最后几滴常常狠压尾巴挤尿。看版上推荐，换了wellness core，两
猫状况不错没拉稀，现在尿团大小排尿姿势正常。
请问，尿尿问题是Royal Canin造成的吗？看成分碳水比重挺高，怎么售价差不多跟
wellness一个价位呀？
Royal Canin babycat36 Ingredients
Chicken meal, rice, corn gluten meal, corn, chicken fat (naturally preserved
with mixed tocopherols, rosemary extract and citric acid), chicken, dried
egg powder, natural chicken flavor, pea fiber, beet pulp, salmon oil, dried
brewers yeast, sodium silico aluminate, potassium chloride, chic... 阅读全帖 |
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p********f
发帖数: 5148 | 19 有人买过这个么？ingredients 非常好的样子。
Deboned Chicken, Deboned Turkey, Chicken Meal, Turkey Meal, Menhaden Fish
Meal (natural source of Omega 3 Fatty Acids), Whole Ground Brown Rice, Whole
Ground Barley, Oatmeal, Peas, Chicken Fat (naturally preserved with Mixed
Tocopherols), Dried Egg, Natural Chicken Flavor, Whole Potatoes, Fish Oil (
natural source of Omega 3 Fatty Acids), Flaxseed (natural source of Omega 3
and 6 Fatty Acids), Whole Carrots, Whole Sweet Potatoes, Cranberries,
Blueberries, Barley Grass... 阅读全帖 |
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z***h
发帖数: 998 | 20 Read following papers:
1.) Wickner RB. [URE3] as an altered URE2 protein - evidence for
a prion analog in Saccharomyces cerevisiae. Science.
264(5158):566-569,1994 Apr 22
This is *the* classic paper that got the whole show on the road
withrespect to prions in yeast.
2.) Edskes HK. Gray VT. Wickner RB. The [URE3] prion is an
aggregated form of Ure2p that can be cured by overexpression of
Ure2p fragments. Proceedings of the National Academy of Sciences of
the United States of America. 96(4):1498- |
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r**r
发帖数: 171 | 21
A Role for Ran-GTP and Crm1 in Blocking Re-Replication.
Yamaguchi R, Newport J.
Department of Biology, University of California, San Diego, 9500 Gilman Drive,
92093, La Jolla, CA, USA
All eukaryotic cells have regulatory mechanisms that limit genomic replication
to a single round each cell cycle. These systems function by blocking
formation of prereplication complexes. The regulatory mechanisms in the yeast
S. cerevisiae have been identified, but these do not appear to be conserved in
metazoan |
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M****e
发帖数: 70 | 22 this is a good paper that tries to dissect the regulatory
mechanism of strict control on DNA replication during cell
cycle. to ensure only one round of DNA replication at each
S phase, yeast S. cerevisiae exploits cdk2 kinase to regulate
functionally reduntant pathways: degradation of Cdc6, export
of MCM from the nucleus, and phosphorylation of ORC. however,
the mechanism in metazoan is unclear while the cellular
targets of cdk2 have not been completely understood yet.
the author (only one besid |
|
t****p
发帖数: 1504 | 23 20世纪70年代初，对DNA聚合酶生化机制的深入了解引申出了一个“复制问题”：由于
DNA聚合酶需要RNA引物来起始DNA复制，线性染色体DNA每复制一轮，都将缩短一个RNA
引物的长度(1)。这意味着细胞需要引入特殊的机制来解决这个“末端复制问题”。而
早在1939年，McClintock报道，在减数分裂后期产生的染色体断裂很容易重新融合起来
，而在紧接着的有丝分裂中，这种染色体“断裂－融合－桥－断裂”的循环将不断继续
(2)。人们从而推测，染色体的自然末端应该不同于一般的DNA断裂末端，它有一个特殊
的结构来避免染色体间的相互融合。
在逐渐明晰了染色体末端特殊结构，即端粒的概念后，Blackburn实验室于1978年第一
次报道，四膜虫（Tetrahymena thermophila）的染色体外线性rDNA的端粒是由重复的5
’-CCCCAA-3’序列组成的(3)。1984年，Blackburn实验室通过将酵母端粒克隆到线性
人工染色体的方法，发现酵母（Saccharomyces cerevisiae）的端粒序列是由不太规则
的TG1-3/C1-3A组成的(4,5)。
在同一篇文章 |
|
q*****d
发帖数: 445 | 24 Plasmid pYD1-CipA1(3)-EGII was obtained by cotransforming G10A1-HR1, G10A1-
HR2,G10A1-HR3, G10A1-HR4, HisG-EGII, and DrdI/NheI-digested pYD1-
CipA1(3) into EBY100. All plasmids were purified from S. cerevisiae and
thentransformed into DH5a, recovered, and confirmed by DNA sequencing.
以上是我在一篇文章里找到的句子，有两个问题不明白，特来请教：
1.这是不是一种多片段克隆的方法，他是把vector和n个要插入的片段纯化之后不用连
接酶直接转化大肠，这种方法成功率如何？多少个片段都可以吗？
2.他的质粒纯化怎么用酵母呢，有什么优点吗？
谢谢大家，因为我有一堆克隆要做，急需要多片段克隆的方法，一个载体有14个片段要
插入！ |
|
h**********r
发帖数: 671 | 25 酵母自己有这样的重组系统，只要片段之间有足够的overlap就可以了。这个叫cloning
by homologous recombination in Saccharomyces cerevisiae
可以看看这个paper，DNA assembler, an in vivo genetic method for rapid
construction of biochemical pathways
P.S
You need a specific strain and plasmid. Go to
http://www.fgsc.net/gpkit.html
and order FGSC 9721 and pRS426.
(FGSC菌株最便宜)
You can follow the knockout protocol to create a long piece of DNA in yeast
through homologous recombination.
http://www.dartmouth.edu/~neurosporagenome/knockout_cassettes. |
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a****o
发帖数: 1786 | 26 今天听Gerald Fink的报告
5% of essential genes in one background of s. cerevisiae is not essential in
S288C background and they are called conditional essentials. |
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s********n
发帖数: 2939 | 27 微生物的模式生物很多啊，S. cerevisiae也是啊，只是说E. coli做的人最多而已。 |
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r**o
发帖数: 212 | 28 细胞培养用的是Sigma的牛胰岛素（insulin solution from bovine pancreas）,但是
现在Sigma缺货，backoeder不知道什么时候才能到货。转用Sigma的合成胰岛素（
recombinant from Saccharomyces cerevisiae）,细胞生长状态变差，出现凋亡状态。
多次从冷冻管活化，生长不到一周又会变差。
求教：不同来源的胰岛素对细胞生长影响那么大么？
有无其他公司售卖的牛胰岛素？
或者用Sigma的牛胰岛素powder替代，唯一的考虑是powder必须用酸性溶液溶解（
dilute acetic or hydrochloric acid, pH 2-3），是否影响细胞生长？原来的牛胰岛
素溶液是现成的溶解在25mM HEPES,pH 8.2里面。
困扰两个月了，求教各位达人了。多谢多谢！ |
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g***j
发帖数: 40861 | 29 一篇文章里说给酵母 S.cerevisiae 转基因
转完以后，分析孢子，发现形状分离比例为2:2
我不懂如何出来的2:2
请教了 |
|
s******i
发帖数: 115 | 30 至少2008年以前，Fan 在西北大学Feinberg School of Medicine做博士后。这些是他
在该实验室的Publication:
Du Z, Park KW, Yu H, Fan Q, Li L.(2008)Newly identified prion linked to the
chromatin-remodeling factor Swi1 in Saccharomyces cerevisiae.Nat Genet. 40(4
):460-5.
Fan Q, Park KW, Du Z, Morano KA, Li L.(2007)The role of Sse1 in the de novo
formation and variant determination of the [PSI+] prion. Genetics. 177(3):
1583-93.
现在在哪就不知道了。 |
|
d*****r
发帖数: 2583 | 31 I think what they mean by genetic network is something described in this one
, you can find the word "genetic network" in the abstract...
And I found your boss' name there, :)
http://www.sciencemag.org/content/327/5964/425.short
The genetic landscape of a cell
Abstract:
A genome-scale genetic interaction map was constructed by examining 5.4
million gene-gene pairs for synthetic genetic interactions, generating
quantitative genetic interaction profiles for ~75% of all genes in the
budding yeast, ... 阅读全帖 |
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a********k
发帖数: 2273 | 32 花了一个小时，深深的鄙视一下自己的无聊行径！！
125 蔡亮 男 1980年11月 复旦大学 生命
科学 2007年12月毕业于[美国]北卡大学 [美国]加州大学旧金山分校 博士后
Cai L, Mostov K. Polarity is destiny. Cell. 2009 Nov 13;139(4):660-2. PubMed
PMID: 19914162; PubMed Central PMCID: PMC2900917.
Cai L, Makhov AM, Schafer DA, Bear JE. Coronin 1B antagonizes cortactin and
remodels Arp2/3-containing actin branches in lamellipodia. Cell. 2008 Sep
5;134(5):828-42. PubMed PMID: 18775315; PubMed Central PMCID: PMC2570342.
Cai L, Makhov AM, Bear JE. F-actin... 阅读全帖 |
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w*******e
发帖数: 63 | 33 你读过snyder前些年的paper吗？
比如
Genes Dev. 1994 May 1;8(9):1087-105.
Large-scale analysis of gene expression, protein localization, and gene
disruption in Saccharomyces cerevisiae.
Burns N, Grimwade B, Ross-Macdonald PB, Choi EY, Finberg K, Roeder GS,
Snyder M.
Source
Department of Biology, Yale University, New Haven, Connecticut 06520-8103.
和
Genome Res. 2004 Oct;14(10A):1975-86.
Large-scale mutagenesis of the yeast genome using a Tn7-derived multipurpose
transposon.
Kumar A, Seringhaus M, Biery MC, Sa... 阅读全帖 |
|
q*****d
发帖数: 445 | 34 我是做合成生物学的，想构建一个新的代谢通路，需要表达很多基因，但是不知道酵母
细胞转化质粒的大小上限是多少？比喻一个30KB的质粒能够通过LiAc方法转化吗？BTW:
我做过最大质粒整合是15KB的。 |
|
h**********r
发帖数: 671 | 35 craig venter同学玩的儿不是更大吗？看看他的protocol。 |
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q*****d
发帖数: 445 | 36 看过他的方法，只是这么大的质粒不知道怎么给转到yeast里面去。 |
|
q*****d
发帖数: 445 | 37 看过他的方法，只是这么大的质粒不知道怎么给转到yeast里面去。 |
|
h**********r
发帖数: 671 | 38 若干有overlap的片段加上plasmid的backbone共转化。利用yeast自身的重组系统，挺
多做酵母的一直这么用。 |
|
|
|
A******r
发帖数: 974 | 41 1.
Mol Gen Genet. 1988 Dec;215(1):10-8.
The detection of mitotic and meiotic aneuploidy in yeast using a gene dosage
selection system.
Whittaker SG, Rockmill BM, Blechl AE, Maloney DH, Resnick MA, Fogel S.
Link:
http://www.springerlink.com/content/l30231v828l8w7q4/fulltext.p
2.
Mutation Research/Genetic Toxicology
Volume 242, Issue 3, November 1990, Pages 231-258
The detection of mitotic and meiotic chromosome gain in the yeast
Saccharomyces cerevisiae: Effects of methyl benzimidazol-2-yl carbam... 阅读全帖 |
|
q*****d
发帖数: 445 | 42 谁有简单易行的方法，需要敲除很多基因，一个个的来还不累死人!这方法到底有木有
啊，目标菌是Saccharomyces cerevisiae.谢谢！ |
|
|
r*****t
发帖数: 4793 | 44 Nature. 2012 Mar 14;483(7390):434-8. doi: 10.1038/nature10895.
Intrinsic coupling of lagging-strand synthesis to chromatin assembly.
Smith DJ, Whitehouse I.
Source
Molecular Biology Program, Memorial Sloan-Kettering Cancer Center, 1275 York
Avenue, New York, New York 10065, USA.
Abstract
Fifty per cent of the genome is discontinuously replicated on the lagging
strand as Okazaki fragments. Eukaryotic Okazaki fragments remain poorly
characterized and, because nucleosomes are rapidly deposited on n... 阅读全帖 |
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l**********1
发帖数: 5204 | 45 if originally omitted below attached figure shown right pathway
then no way can bet below report:
Levy SF et al.(2012)
Bet hedging in yeast by heterogeneous, age-correlated expression of a stress
protectant.
PLoS Biol. 10: e1001325.
Abstract
Genetically identical cells grown in the same culture display striking cell-
to-cell heterogeneity in gene expression and other traits. A crucial
challenge is to understand how much of this heterogeneity reflects the noise
tolerance of a robust system and ho... 阅读全帖 |
|
l**********1
发帖数: 5204 | 46 sure
add one recent paper
Levy S at al. (2011)
The competitive advantage of a dual-transporter system.
Science. 334: 1408-12.
Abstract
Cells use transporters of different affinities to regulate nutrient influx.
When nutrients are depleted, low-affinity transporters are replaced by high-
affinity ones. High-affinity transporters are helpful when concentrations of
nutrients are low, but the advantage of reducing their abundance when
nutrients are abundant is less clear. When we eliminated such red... 阅读全帖 |
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l**********1
发帖数: 5204 | 47 Standard error of mean (SEM)
Dynamics of cortical actin in budding yeast Saccharomyces cerevisiae
Dissertation zur Erlangung des Doktorgrades
Der Fakultät für Chemie und Pharmazie
der Ludwig-Maximilian-Universität München (2010).
web link:
Http://edoc.ub.uni-muenchen.de/11423/1/Yu_Haochen.pdf
or
Higgs PG
Error thresholds and stationary mutant distributions in multilocus diploid
genetics models (1994)
Genet. Res. Cambridge 63: 63-78.
web link
//siba.unipv.it/fisica/articoli/G/GeneticalR... 阅读全帖 |
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h**********r
发帖数: 671 | 48 Title:
The red/white colony color assay in the yeast Saccharomyces cerevisiae:
epistatic growth advantage of white ade8-18, ade2 cells over red ade2 cells
Current Genetics
December 1996, Volume 30, Issue 6, pp 485-492
my email is m******[email protected], Thank you so much! |
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r*********g
发帖数: 1160 | 49 谢谢回复，请问你们震动60秒一般是多大体积的culture？产热多么？
另外除了s.cerevisiae，还用于别的organisms么？ |
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