k***g
发帖数: 4904 | 1 配working solution用啥media啊，manual上只是写“Dilute the stock solution to
a final working concentration of 0.5-25 µM in serum-free medium. Avoid
amine- and thiol- containing buffers.”像E.coli这种在LB broth里面养的细菌，
应该用什么media配这种working solution呢？谢谢！ |
|
k***g
发帖数: 4904 | 2 E coli ATCC25922，suspended in 4°C PBS，放一两天活力会下降多少？大部分能撑
一个星期不死吗？
如果我想做一个星期实验（或者说3天左右吧），不反复配细胞浓度，放在4°C可以不？是不是如
果放在media里面，会活的长些，但是有可能会变浓，在PBS里不会变浓，但是可能早点死翘翘？ |
|
m****M
发帖数: 360 | 3 Depends on the strain, but one week is normally ok for most E.coli strains. |
|
h**********r
发帖数: 671 | 4 想用Wanner Lambda Red Gene Disruption系统敲除E. coli的6个基因，别人报道过了
，没有问题。
一个一个的敲费时啊，能不能同时转化两个PCR产物，分别用Km和Cm的抗性，筛选Km Cm
的抗性，然后切断marker,再做下一次的两个基因的敲除。
求建议！有包子！
多谢！ |
|
l**********1
发帖数: 5204 | 5 老大或你老板 能开发个Array knock out tool that can parallel 敲除任意N (N>2) N个基因 at E coli
next Yeast, fly, zebrafish, especially on Rat Mice Human at universal biomedical reactions pathway middle
point. then 这里的千老就能成墙街矿工或湾区龟公的生路喽
而你和你老板可以炸药奖或被至少被那个奖提名
of course, might should be done at Biosafety level P4 areas. -- not joke, Sure!
//en.wikipedia.org/wiki/Biosafety_level#Biosafety_level.C2.A04
Cm |
|
h**********r
发帖数: 671 | 6 我老板不管我，自己瞎折腾。
给PNAS的原作者写了封信问这事儿，人家懒得搭理我。
不过你这推论也太离谱了，从e. coli到Rat Mice Human，从苦逼到炸药奖或被至少被
那个奖提名。
算了，我还是老老实实的挨个儿敲吧。
多谢！ |
|
x********u
发帖数: 430 | 7 I have finished a dozen of deletions in E coli and based on my experience
there is no way you can simultaneously knockout two genes using this Red
recombination method. The chances are very low if your PCR fragment has only
40-50 homologous region with your target gene. Without removing the
introduced FRT site could cause site-unspecific integration and the second
deletion could happen nowhere but your FRT site. |
|
s*******s
发帖数: 623 | 8 streptomyces上的应用实际上并不是直接在streptomyces中用的，用E. coli穿梭一下
而已 |
|
z**m
发帖数: 50 | 9 请教版上有没有了解E.coli beta galactosidase结构功能的大牛？目前准备个试验，
需要构建无活性的beta galactosidase突变体做control，因为不是生物背景，查了很
多文献，还是没有头绪，不知道哪些位点的突变或者缺失可以完全abolish beta
galactosidase的活性但是又不影响酶的正常tetramer结构的。有没有哪位大牛可以指
点一下或者提供一两篇类似的文献报道？包子答谢！谢谢！ |
|
w*****t
发帖数: 179 | 10 My co-worker asked me this question, what is minimal E.coli extract, I have
no idea what this is. Is anybody here can help me? Is there any paper to
recommend me to read about this? Thanks a lot. |
|
b******y
发帖数: 627 | 11 More context help nail it down.
In NMR field, we use minimal medium to culture E. coli. But not sure whether
it is related to your question. |
|
h**********r
发帖数: 671 | 12 在给一个真菌做cre/lxop的knockout系统，想在一个质粒中装好cre和marker，两端是
loxp。cre克隆在真菌的诱导表达的启动子下游。cassette是这样子的：loxp-marker-
Promoter-cre-terminator-loxp。我是从AT克隆开始做的,pgem-t easy vector.问题是
最后一步怎么都装不上去。大小好像回到了最开始的pgem-t easy的载体上去了。初步
估计这个promoter在E. coli中也有表达，然后自己就把中间的那部分切下去了。试了
好几次，同样的情况。当然还没有sequence.请问大家遇到这种情况怎么办？ |
|
h**********r
发帖数: 671 | 13 Title:
Flux analysis and control of the central metabolic pathways in Escherichia
coli
FEMS Microbiology Reviews
Volume 19, Issue 2, pages 85–116, December 1996
http://www.ncbi.nlm.nih.gov/pubmed/8988566
My email is m******[email protected]
One Baozi will be sent after getting the paper.
Thanks! |
|
jh
发帖数: 23 | 14 实验室-80C坏了，正在维修，请问能否把E. coli glycerol stock移到-20度暂时放1，
2天？
谢谢！ |
|
e****s
发帖数: 1125 | 15 LS太武断了。
没问题。E.coli 很tough的。 |
|
s******r
发帖数: 1245 | 16 2a难道不是真核，你肯定在e.coli里能用？ |
|
发帖数: 1 | 17 想用lamda red在BL21中敲一个基因，按照protocol，把pKD46转入BL21，很奇怪的是不
管用化转还是电转，涂板后都没有长出来，但是转其他的质粒都没有问题。对pKD46跑
胶和单酶切都没有问题，按照protocol所有incubation的温度都是30度。同时还试过E.
coli 另外的菌株BW25113也是出现同样问题。有人遇到过质粒转成这样的情况吗？多谢
！ |
|
H*******i
发帖数: 196 | 18 “试过E.coli 另外的菌株BW25113也是出现同样问题”
质粒问题呗。 当然最好看看你们的培养箱的30°确认下没问题，不能确认就放室温
（25°）长，也应该能长出来。
另外多说一句，这个pKD46构建比较早了，KO效率不如一些新构建的。
还有听一些人说过BL21 用这个方法KO好像很难？ 不过这个我没验证过。 |
|
b*****o
发帖数: 6080 | 19 大家谁能给科普一下E. Coli来源，感染途径，受影响最大的器官，以及治疗方法？ |
|
b*****o
发帖数: 6080 | 20 大家谁能给科普一下E. Coli来源，感染途径，受影响最大的器官，以及治疗方法？ |
|
发帖数: 1 | 21 ◇◇新语丝(www.xys.org)(newxys.com)(xys10.dxiong.com)◇◇
举报南京大学生命学院院长华子春学术不端
方先生您好！
我们反映南京大学生命学院院长、长江、杰青、 国家科技进步奖获得者、
国重主任华子春长期存在学术不端问题。 附件是我们发现的第一批四组共9篇文
章。
1. 2000-C和2000-E两篇文章皆为英文稿， 分布发表于南京大学学报和
Protein Expression and Purification。华教授是通讯作者。 南京大学学报的
内容系于后者的文字上做了删掉，无文字改写。 例如：
前者的第一、二段：
Human cardiac-specific homeobox protein (hCsx2 or Nkx2.5) encodes
a homeobox transcription factor of 323 amino acids containing six
cysteines and is composed of three domains: the TN-domain, the
homeobox- domain, and t... 阅读全帖 |
|
S*****P
发帖数: 323 | 22 http://news.sciencemag.org/scienceinsider/2011/06/sequence-yiel
DNA Sequence Yields Clues to Germany's 'Super Toxic' E. coli outbreak
by Martin Enserink on 2 June 2011, 5:50 PM |
Just from the high number of deaths and severe cases, scientists and public
health experts battling Germany's massive E. coli outbreak knew they were up
against something unusual. Now, early results from sequencing projects of
the enterohemorrhagic Escherichia coli (EHEC) strain appear to confirm that
a never-before-see... 阅读全帖 |
|
s********n
发帖数: 2939 | 23 Host strain Genotype
BL21-CodonPlus(DE3)-RIPL strain E. coli B F– ompT hsdS(rB
– mB–) dcm+ Tetr gal λ(DE3) endA Hte [argU proL Camr] [argU ileY leuW
Strep/Specr]
BL21-CodonPlus-RIL strain3 E. coli B F– ompT hsdS(rB
– mB–) dcm+ Tetr gal endA Hte [argU ileY leuW Camr]
BL21-CodonPlus(DE3)-RIL strain3 E. coli B F– ompT hsdS(rB
– mB–) dcm+ Tetr gal λ(DE3) endA Hte [argU ileY leuW Camr]
BL21-CodonPlus(DE3)-RIL-X strain3 E. coli B F– ompT hsdS(rB
– mB–) dcm+ Tetr gal λ(DE3) endA metA::Tn5(kanr) Hte [ar... 阅读全帖 |
|
C*******e
发帖数: 4348 | 24 谢谢这么详尽的trouble shooting
1. 限制性内切酶切E.coli plasmid显示，根据我们手上有的序列信息，如果只切E.
coli源的那
一半，酶切图谱就是正确的；如果在细菌X的origin那一段序列也切，酶切图谱就不能
完全吻合。所
以现在已经放弃用那个E.coli plasmid。
2.这个E.coli plasmid，同一管样品，别的人做PCR，还有ligation，酶切等等（都在E
.coli
源的那一半）是完全没有问题的，所以排除E.coli plasmid不够纯的问题。
3.同样的PCR master mix，用不同模板、引物，是可以PCR没有问题的，所以和试剂没
有关系。
4.引物序列等等已经确认过很多次；不过我没有确认到底有多少DNA在里面，谢谢提醒
，回头会查查
看。
5.要扩增的序列GC%是60%，不算特别高。PCR enhancer等等也试过，没有帮助。
6.我还提到如果不加DNA聚合酶的话加样孔里就没有东西；后来试过酚氯仿提取、乙醇
沉淀，再跑胶
只有一点smear，所以现在确定那是蛋白+核酸在孔里；为什么同样的master mix做别的
P |
|
n*********m
发帖数: 38 | 25 Look his publication
Conversion of D-ribulose 5-phosphate to D-xylulose 5-phosphate: new insights
from structural and biochemical studies on human RPE.
Liang W, Ouyang S, Shaw N, Joachimiak A, Zhang R, Liu ZJ.
FASEB J. 2011 Feb;25(2):497-504. Epub 2010 Oct 5.
PMID:
20923965
[PubMed - indexed for MEDLINE]
Related citations
2.
Structural basis for the inhibition of human 5,10-methenyltetrahydrofolate
synthetase by N10-substituted folate analogues.
Wu D, Li Y, Song G, Cheng C, Zhang R, Joac... 阅读全帖 |
|
c********n
发帖数: 225 | 26 对于韩春雨NBT文章无法重复的问题
需要从Figure 1 开始一步步看
作者的contributions
1. C.H. conceived the study and designed the experiments.
文章 “Story” “Idea” 的设计与发起人
2. X.Z.S. provided intellectual advice on the project and experimental
design.
实验设计， “One guide-faithful” -Figure 2d？
3. C.H. performed mammalian genome editing.
（Figure 4,5）
文章里让科研界最感兴趣的结果，也是无法被重复验证的主要问题
4. F.G. performed the BLAST search and the in vitro cleavage experiments.
（Figure 1, supplementary figure 2）
大肠杆菌E coli 表达的蛋白
NgAgo 基因编辑功能的初步 in vitro 数据支持
注... 阅读全帖 |
|
c********n
发帖数: 225 | 27 【再贴一次】
对于韩春雨NBT文章无法重复的问题
需要从Figure 1 开始一步步看
作者的contributions
1. C.H. conceived the study and designed the experiments.
文章 “Story” “Idea” 的设计与发起人
2. X.Z.S. provided intellectual advice on the project and experimental
design.
实验设计， “One guide-faithful” -Figure 2d？
3. C.H. performed mammalian genome editing.
（Figure 4,5）
文章里让科研界最感兴趣的结果，也是无法被重复验证的主要问题
4. F.G. performed the BLAST search and the in vitro cleavage experiments.
（Figure 1, Figure 2？，Figure 3a，supplementary figure 2）
大肠杆菌E coli 表达的蛋白
NgAg... 阅读全帖 |
|
w*****t
发帖数: 179 | 28 您好，不知道我的背景能否审到这片文章。我曾经审过一些小杂志，希望能够对我争取
这次机会有所帮助。详细的CV和研究背景如下。非常感谢您的考虑。我的工作邮箱：
z****[email protected]
好像CV在这里粘贴很难看，不好意思。前一段时我的CV，后一段我的较详细的研究经历
。如有需要我更详细的背景情况，请跟我发信。
1
CURRICULUM VITAE
Full Name: Zhimin Wang
Address: Allegheny-Singer Research Institute, West Penn Allegheny, Health
System, South Tower, 320 East North Avenue, Pittsburgh, PA 15212-4772
Phone: 412-359-6153 (Office)
412-737-3777 (Cell)
E-mail: z****[email protected] w************[email protected]
EDUCATION
September, 2001- May, 2005 Ph.D., Molecular M... 阅读全帖 |
|
w*****t
发帖数: 179 | 29 hello, thank you for providing the information. Please find my CV below and
my research experience. I have been revewing for over 10 journals for 14
times. My majour of PH.D is microbiology from Wuhan university. I have
published several papers concering virology and bacterial infection. So far
my work also foucse on salivary gland biofilm formation under LL37 and K4s4
treatment. If you need further information, please let me know. My email
address: z****[email protected]
Thank you for your time and c... 阅读全帖 |
|
s***e
发帖数: 911 | 30 Sunny Xie最近有个工作,是在bacterial level理解epigenetics. 方法是在Lac genes上
标记一个YFP， 然后用显微镜定量每个E.Coli的荧光强度. 从一个single e.coli开始,
跟踪观察其后代. 因为Lac repressor的关系, YFP表达是比较低水平的. 随着分裂继
续, 一个pheno type出现，这些e.coli具有高表达的YFP. 这个特征可以遗传下去.
解释: e.coli里只有几个Lac Repressor. 一个reporessor有两个DNA binding sites.
热力学上具有很小的机会让这个repressor完全离开DNA. 一旦完全离开, 因为inducer
会compete bind 到repressor上, 则rebinding的kinetics会非常慢. 于是在一个相当
长的时间内, 这些e.coli的特性会遗传到下一代.他强调的是, 至少对他研究的这个
model system来说, epigenetics无非是一个小概率偶发事件, 并且远离热力学平衡态
的结
果.
我这也是报告会上听的。不 |
|
x****6
发帖数: 4339 | 31 说先怎么定义 从e coli进化出酵母？
如果我从E COLI进化出，一个e coli生存在 另一个e coli里面，给它补充能量来源，
算不算？
这个给我10年，我可以整出来。
不过就算整出来，也没有卵用。浪费钱 |
|
W*******n
发帖数: 2762 | 32 Scientist/ Senior of Molecular Biology
Challenges
You will be involved in cross-functional project teams with colleagues from
many different scientific backgrounds both in China and at our head quarters
in Denmark. You will contribute to several early stage projects with
responsibility for designing expression systems and generating recombinant
proteins preferentially from E. coli, but some experience within mammalian
expression is also considered valuable. You will also be responsible for
proce... 阅读全帖 |
|
W*******n
发帖数: 2762 | 33 Scientist/ Senior of Molecular Biology
Challenges
You will be involved in cross-functional project teams with colleagues from
many different scientific backgrounds both in China and at our head quarters
in Denmark. You will contribute to several early stage projects with
responsibility for designing expression systems and generating recombinant
proteins preferentially from E. coli, but some experience within mammalian
expression is also considered valuable. You will also be responsible for
proce... 阅读全帖 |
|
C*******e
发帖数: 4348 | 34 乘着这会儿人气高，求教一个PCR中遇到的问题哈，说实在的这是第一次遇到这么诡异的PCR问题，
这么多年下来大大小小各式各样的PCR都做过，扩增大于10kb都没有遇到过不知道如何trouble
shooting的时候。
最近从某个E.coli plasmid里PCR一个非E.coli的origin X(这个plasmid既有Ecoli的
origin又有另外某个细菌X的origin X，既可以在E.coli里维持也可以在细菌X里维持)。这个
origin X有3.6kb长，照理说用plasmid做模板，3.6kb也不算多长，应该不会太麻烦；可是做下
来跑胶就是干干净净，没有条带，没有primer dimer，没有非特异产物，什么都没有（当然如果模
板plasmid加多一点，能看到模板）；annealing温度梯度，还有别的optimize都做了，还是一
样；还有过用linear plasmid做模板，还是一样。
另外还设计了这个3.6kb区域的5个sequencing primer，因为一直P不出来，就有点怀疑这个
E.coli plasmid，因为是别的实验室给我们的，送去测序以后没有 |
|
b******r
发帖数: 111 | 35 I am doing a knockin project. I have done constructs which contain Neo and
HSV-TK for selection in ES cells.
How do I know Neo and HSV-TK in my plasmid work? Specificly,
Do Neo and HSV-TK genes in my plasmids express their proteins in E.Coli? I
tested in this way: grow 4 tubes
of E.Coli, when adding G418, E.Coli can't grow; When adding gancyclovir, E.
Cori grow well. That means Neo
and HSV-TK genes of my plasmids don't express in E.Coli, right? Then why
do Neo and HSV-TK express in
ES cells? ... 阅读全帖 |
|
g***j
发帖数: 40861 | 36 please email to s*****[email protected]
Thank you very much!
1: Erickson MC, Liao J, Payton AS, Riley DG, Webb CC, Davey LE, Kimbrel S,
Ma L,
Zhang G, Flitcroft I, Doyle MP, Beuchat LR. Preharvest internalization of
Escherichia coli O157:H7 into lettuce leaves, as affected by insect and
physical
damage. J Food Prot. 2010 Oct;73(10):1809-16. PubMed PMID: 21067668.
2: Erickson MC, Webb CC, Diaz-Perez JC, Phatak SC, Silvoy JJ, Davey L,
Payton AS,
Liao J, Ma L, Doyle MP. Surface and internalized Escherich... 阅读全帖 |
|
a*****x
发帖数: 901 | 37 ****************************************************************************
***
人物
1． Wilbrandt的核心观点在于Carrier通过拓扑结构的改变, 使得其中的离子只能
向一个方向运动. 这一点虽然以前也有很多人提到过, 但是Wilbrandt应该是第一个做
出另人信服的实验的人. 这个模型后来被简化为我描述的双门模型, 其本质是一样的.
2． 第三章轻松一下, 介绍几个主角, 没有reference....
先来介绍我们的第一位主角, Christopher Miller. 这是一个人的名字. 这位仁兄在
ion channel学界是一位绝对的“大牌“. 当他出现在任何一个neuroscience meeting
或者biophycis meeting的时候, 几乎都是所谓的keynote speaker. 他的成就太多, 无
法一一列举. 比如, 他是第一批记录到单通道电流的人之一. 什么是单通道电流呢? 因
为这个以后要用到, 所以我在这里讲一下.
所谓单通道电流, 就是指通过... 阅读全帖 |
|
a****k
发帖数: 1130 | 38 J Bioenerg Biomembr. 2002 Dec;34(6):423-31.
Expression and characterization of recombinant human cytochrome c in E. coli.
Jeng WY, Chen CY, Chang HC, Chuang WJ.
Source
Department of Biochemistry, National Cheng Kung University College of
Medicine, Tainan 701, Taiwan.
Abstract
Cytochrome c is a heme protein involved in electron transfer, cell apoptosis
, and diseases associated with oxidative stress. Here we expressed human
cytochrome c in E. coli and purified it to homogeneity with a yield of 10... 阅读全帖 |
|
d******n
发帖数: 29 | 39 1.
Towards large scale methods for the selective release of periplasmic human
cystatin C from E. coli
Biotechnology Techniques, Volume 9, Number 3 (1995), 179-184
2.
Optimization of an osmotic shock procedure for isolation of a protein
product expressed in E. coli.
Biotechnol Prog. 2003 Sep-Oct;19(5):1541-6.
3.
Optimization of BLyS production and purification from Escherichia coli.
Protein Expr Purif 39:237-246.
4.
High-level accumulation of a recombinant antibody fragment in the periplasm
of Es... 阅读全帖 |
|
d******n
发帖数: 29 | 40 1.
Towards large scale methods for the selective release of periplasmic human
cystatin C from E. coli
Biotechnology Techniques, Volume 9, Number 3 (1995), 179-184
2.
Optimization of an osmotic shock procedure for isolation of a protein
product expressed in E. coli.
Biotechnol Prog. 2003 Sep-Oct;19(5):1541-6.
3.
Optimization of BLyS production and purification from Escherichia coli.
Protein Expr Purif 39:237-246.
4.
High-level accumulation of a recombinant antibody fragment in the periplasm
of Es... 阅读全帖 |
|
h**********r
发帖数: 671 | 41 直接把你的AA序列发给公司，像genscript.然后就可以拿到quote,保含cia,gc content
， mRNA二级结构，这一步是free的。
再决定要合成不。
avoid after initiating met: vla, lys, leu, tyr, arg, phe ,trp
best codons in e. coli: ala (gcg), gly (ggc, ggu)
也可以看看老paper:
Expression of Mammalian Cytochromes P450 in Yeast
Expression of Eukaryotic Cytochromes P450 in E. coli
虽然讲的是p450,有些策略还是可以借鉴的。
可以看下新的paper:
Design Parameters to Control Synthetic Gene Expression in Escherichia coli
Kudla, G. et al. Coding-sequence determinants of gene expression in
Escheri... 阅读全帖 |
|
d******n
发帖数: 29 | 42 1.
Towards large scale methods for the selective release of periplasmic human
cystatin C from E. coli
Biotechnology Techniques, Volume 9, Number 3 (1995), 179-184
2.
Optimization of an osmotic shock procedure for isolation of a protein
product expressed in E. coli.
Biotechnol Prog. 2003 Sep-Oct;19(5):1541-6.
3.
Optimization of BLyS production and purification from Escherichia coli.
Protein Expr Purif 39:237-246.
thanks a lot!
4.
High-level accumulation of a recombinant antibody fragment in the p... 阅读全帖 |
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h*******e
发帖数: 1377 | 43 如果 m n 中有一个小于３０的话
for(int rowI = 0; rowI < rowNum; ++ rowI)
for(int dpI = 0; dpI < 1 << colNum; ++ dpI)
for(int colI = 0; colI < colNum; ++ colI)
{
//然后自己转移状态就行了。
３ 种情况 １ 本格占了 。
２ 本格没占 右边突出
３ 本格没占 下边突出
然后相加可能的各种情况就行了。
}
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h*******e
发帖数: 1377 | 44 如果 m n 中有一个小于３０的话
for(int rowI = 0; rowI < rowNum; ++ rowI)
for(int dpI = 0; dpI < 1 << colNum; ++ dpI)
for(int colI = 0; colI < colNum; ++ colI)
{
//然后自己转移状态就行了。
３ 种情况 １ 本格占了 。
２ 本格没占 右边突出
３ 本格没占 下边突出
然后相加可能的各种情况就行了。
}
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M******3
发帖数: 7 | 45 ● China Opportunity _Principal Scientist or Sr. Scientist
We are retained by a global Pharma company in China to find Principal
Scientist or Sr. Scientists in Molecular Biology
m*********[email protected] (Monday to Friday)
m***********[email protected] (Weekend)
POSITION DESCRIPTION
Sr._Scientis/Principal_Scientist_Molecular_Biology
Summary of Key Responsibilities:
• Develop effective microbial host/vector systems using recombinant
DNA technology, including:
identification of expression lim... 阅读全帖 |
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w*****t
发帖数: 179 | 46 Thank you for providing the review opportunity, I have been working in the
molecular biology for over 15 years and published some papers in journals
like Journal of Cell Biology, Journal of General Virology et al., . Also I
have reviewed over 20 manuscripts for 10 journals. Hopefully these
experiences will help me to catch this opportunity. If you need further
detail information regarding my qualification, please let me know (zwang1@
wpahs.org). Please find my CV below. Thanks a lot.
Zhimin Wang... 阅读全帖 |
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c*******3
发帖数: 98 | 47 I would like to review ZnO growth manuscript.Thanks!
My CV is as follows:
JINGBO CHANG
Laboratory of Nanotech. for Sustainable Energy and Environment University of
Wisconsin-Milwaukee, 3200 North Cramer Street Milwaukee, WI 53211, USA
www.researchgate.net/profile/Jingbo_Chang/publications/
PERSONAL DETAILS
Date of Birth: 28th Sep. 1979 Gender: Male
Nationality: China Marital Status: Married
Tel. +1- 414- 688-1383 E-mail: c*****[email protected]
EDUCATIONS
☼ Post doc, major in Mechanical Engineerin... 阅读全帖 |
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s***e
发帖数: 911 | 48 DNA topoisomerases are enzymes that catalyze the passage of individual DNA
strands or double helices through one another. These reactions are often
manifested in interconversions between topological isomers of DNA rings;
hence the name of the enzymes...
There are three enzymes in this group: E.coli DNA topoisomerase I,
mouse DNA topoisomerase I, and E.coli topoisomerase II, respectively,
representing 3 distinct subfamilies of topoisomerases.
Both E.coli DNA topoisomerase I and mouse DNA topoisom |
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z****g
发帖数: 972 | 49 要求：
1.来源于Col-0的幼苗
2.经过了nomalization,减少了abundant transcripts,比如actin, tubulin等
3.host为普通E.coli,比如W3110或DH5a
4.所在质粒需为E.coli compatible的质粒，本身带有RBS (ribosome binding site)
5.可以受IPTG诱导直接在E.coli中表达
自己google了一堆，没找到合适的。如果板上哪位知道哪里可以购买，请pm |
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x*********a
发帖数: 906 | 50 E. coli host genotype has to be lamda+?
lamda- strains can not express T7 promoter-direced gene from plasmid?
Any other prophage on E. coli K12 has T7 RNA polymerase?
How about SP6 promoter? what kind of genotype can not express SP6 promoter ?
Can this genotype E. coli K12 ∆(araD-araB)567, ∆lacZ4787(::rrnB-
3), lambda-, rph-1, ∆(rhaD-rhaB)568, hsdR514 express SP6 or T7
promoter-driven gene?
Thanks a lot. |
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