l**********1
发帖数: 5204 | 1 GST-A.B co-expression pls let IPTG and the cultures were maintained at 4°C
for 48-72 h then do that induction
reference:
PMID 23525091
Production of soluble eukaryotic recombinant proteins in E. coli is favoured
in early log-phase cultures induced at low temperature. (2012).
Springerplus. 2:89.
FINDINGS:
A high yield of active soluble proteins was obtained by combining early-log
phase cultures and low temperatures for protein induction. When IPTG was
added at OD600 = 0.1 and cultures... 阅读全帖 |
|
l***d
发帖数: 1798 | 2 我给你总结一下这个文章的大意，
文科生
这个文章是说prion 和ad有些共性
1.虽然这两个病设计不同的蛋白和多肽，但是都能misfold and aggregate成结构相似的
amyloid fibrils with cross-beta structures.
2. 同时misfold的过程，经常参与一些cu的结合（其实不仅仅是cu, zn, ca在不同neur
odegenerative dieseases都会参与）
文科生，露怯了
这篇文章，有个毛证据是说
“很多ad病人是吃疯牛得病的” |
|
s******s
发帖数: 13035 | 3 我不是说了么。misfold了就容易降解。
我只能说很多lof的mutant应该会misfold, 至于你的mutant到底是啥，只有你知道
我不是这行的，knowledge就这么多了
proteasome |
|
r*******g
发帖数: 32828 | 4 我的2011年版找工简历
王利民
2014年8月16日
（现注：当时，未列出2010年在Fox Chase Cancer Center的经历；没列出2005年在
Memorial Sloan-Kettering Cancer Center的经历；没列出2006年在陕西师范大学、
2009年在江南大学任教的经历。）
Career Summary:
PhD degree on Chemistry and post-doctoral research on protein
structure in the USA; Teaching (assistance) experience in the USA and
China; Customer service experience in the USA.
Work/ Research:
Jan 2006 - : various work in the USA, China, and home.
Sept. 2003- June 2005: New York State Department of Health
Post-doctorate
Re... 阅读全帖 |
|
b****g
发帖数: 1204 | 5 【 以下文字转载自 WaterWorld 讨论区 】
发信人: buydig (digbuy), 信区: WaterWorld
标 题: 这王利民是混的最惨的高考状元吗？
发信站: BBS 未名空间站 (Tue Aug 19 22:40:39 2014, 美东)
Honor:
Highest score in the 1991' National College Entrance Exam in Hunan
Province, China
2011年版找工简历
王利民
Career Summary:
PhD degree on Chemistry and post-doctoral research on protein
structure in the USA; Teaching (assistance) experience in the USA and
China; Customer service experience in the USA.
Work/ Research:
Jan 2006 - : various work in the USA, China, and ho... 阅读全帖 |
|
c****y
发帖数: 101 | 6 发信人: WangLimin (王利民), 信区: WaterWorld
标 题: 我的2011年版找工简历
发信站: BBS 未名空间站 (Sat Aug 16 09:18:20 2014, 美东)
我的2011年版找工简历
王利民
2014年8月16日
（现注：当时，未列出2010年在Fox Chase Cancer Center的经历；没列出2005年在
Memorial Sloan-Kettering Cancer Center的经历；没列出2006年在陕西师范大学、
2009年在江南大学生物工程学院任教的经历。）
Career Summary:
PhD degree on Chemistry and post-doctoral research on protein
structure in the USA; Teaching (assistance) experience in the USA and
China; Customer service experience in the USA.
Work/ Research:
Jan 2006 - : various ... 阅读全帖 |
|
b*****z
发帖数: 163 | 7 哪位大侠帮忙查下
VIN:
1FMDU32E1XZC07501
Email:
m********[email protected]
非常感谢！
切查询都是依靠其他志愿帮助的id提供Carfax资料。请寻求帮助的同学谅解。如果几天
内甚至永远没有人为您提供帮助，也是正常。我想是不是可以设置一个时间，比如3天
。如果没有答复就可能是没有人愿意提供帮助，发帖者本人要自行删旧帖，顶新贴。另
外，我善意的提醒大家，对一个不付费的服务就不要期望太高，否则就该自己购买服务。 |
|
b*****z
发帖数: 163 | 8 哪位大侠帮忙查下
VIN:
1FMDU32E1XZC07501
Email:
m********[email protected]
非常感谢！
切查询都是依靠其他志愿帮助的id提供Carfax资料。请寻求帮助的同学谅解。如果几天
内甚至永远没有人为您提供帮助，也是正常。我想是不是可以设置一个时间，比如3天
。如果没有答复就可能是没有人愿意提供帮助，发帖者本人要自行删旧帖，顶新贴。另
外，我善意的提醒大家，对一个不付费的服务就不要期望太高，否则就该自己购买服务。 |
|
W*******n
发帖数: 2762 | 9 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 |
|
b****g
发帖数: 1204 | 10 【 以下文字转载自 WaterWorld 讨论区 】
发信人: buydig (digbuy), 信区: WaterWorld
标 题: 这王利民是混的最惨的高考状元吗？
发信站: BBS 未名空间站 (Tue Aug 19 22:40:39 2014, 美东)
Honor:
Highest score in the 1991' National College Entrance Exam in Hunan
Province, China
2011年版找工简历
王利民
Career Summary:
PhD degree on Chemistry and post-doctoral research on protein
structure in the USA; Teaching (assistance) experience in the USA and
China; Customer service experience in the USA.
Work/ Research:
Jan 2006 - : various work in the USA, China, and ho... 阅读全帖 |
|
l**********1
发帖数: 5204 | 11 楼主一个lab this fiscal year can get 7 M usd for next 3 to 5 Ys? if not DOM
relavant idea, less convincing...
if included collabrative labs, it is normal,
for example: 8 labs one
EU Research Projects
MULTISYN
Multimodal Imaging of rare Synucleinopathies
From 2013-10-01 to 2018-09-30 | MULTISYN website
Project details
Despite remarkable progress in the understanding of the molecular
pathogenesis of neurodegenerative diseases (NDD), no causative or disease-
modifying treatment is currently available... 阅读全帖 |
|
M******3
发帖数: 7 | 12 ● 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 |
|
W*******n
发帖数: 2762 | 13 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******8
发帖数: 62 | 15 四岁半的女儿非常淘气，妈妈对她比较严厉，但是爸爸对她很宠，所以她经常欺负老实
巴交的爸爸，把爸爸耍得团团转。
一天早上醒来，爸爸发现女儿的鼻子上有点鼻血，就说：“你鼻子上有鼻血，我去拿张
paper towel擦擦。”
等爸爸拿了张paper towel来，要擦的时候，女儿说：No， I need a towel.于是爸爸
就去拿她常用的红色小毛巾。
再要擦的时候，女儿说：No， I need a wet towel. 爸爸又回头去把毛巾打湿，心想
这下好了吧。
女儿说： No， I need a super wet towel. 老实的爸爸又去了次bathroom，出来说：
“快， super wet towel 来来，还在滴水。”
女儿把头一扭，说：“No， I don't want that."
我听了又好气又好笑，于是黑脸妈妈出来了，说：“你不是要做five star girl吗？刚
才那样子只能拿5 gray star了，就是只有zero gold star了。”
女儿听完，立刻从爸爸手里接过毛巾，自己把鼻血擦干净了。爸爸私下对我说：“要是
你，肯定不会给她跑四趟。”
中文儿... 阅读全帖 |
|
|
b*****z
发帖数: 163 | 17 转帐完成
转给用户:kbrhouston,现金(伪币):100,收取手续费:1.00
email: m********[email protected]
谢谢 |
|
b*****z
发帖数: 163 | 18 Big Congratulations!
Could you plz send a copy of your application docs to me?
email: m********[email protected]
Thank you very much! |
|
b*****z
发帖数: 163 | 19 Could you plz also send me a copy?
email: m********[email protected]
Thanks |
|
|
t****l
发帖数: 445 | 21 我的基本情况：PhD in Theoretical biophysics, 2008. Currently, I have been in
Postdocoral position for 4 years. I am focusing on developing statistical
modeling of protein folding, protein misfolding (such as amyloid fiber
formation), aggregation of bacterial cell and polymer degradation.
If by any chance, I would like to have some 审稿机会 for my application of
green card. Please send email to me n**********[email protected].
多谢。 |
|
t****l
发帖数: 445 | 22 My background:
PhD in Theoretical biophysics, I am focusing on developing statistical
modeling of protein folding, protein misfolding (such as amyloid fiber
formation), aggregation of bacterial cell and polymer degradation.
If by any chance, I would like to have some 审稿机会 for my application of
green card.
多谢。 |
|
t****l
发帖数: 445 | 23 My background:
PhD in Theoretical biophysics, I am focusing on developing statistical
modeling of protein folding, protein misfolding (such as amyloid fiber
formation), aggregation of bacterial cell and polymer degradation.
If by any chance, I would like to have some 审稿机会 for my application of
green card.
多谢。 |
|
W*******n
发帖数: 4140 | 24 我的2011年版找工简历
王利民
2014年8月16日
（现注：当时，未列出2010年在Fox Chase Cancer Center的经历；没列出2005年在
Memorial Sloan-Kettering Cancer Center的经历；没列出2006年在陕西师范大学、
2009年在江南大学任教的经历。）
Career Summary:
PhD degree on Chemistry and post-doctoral research on protein
structure in the USA; Teaching (assistance) experience in the USA and
China; Customer service experience in the USA.
Work/ Research:
Jan 2006 - : various work in the USA, China, and home.
Sept. 2003- June 2005: New York State Department of Health
Post-doctorate
... 阅读全帖 |
|
f*****g
发帖数: 15860 | 25 lol, this is why i like to play some live games once in a while, it's
totally different experience... no stats to check, no misfold button, timer
is there but not as strict as online, dirty play "i can show you one card
for $100", trash talk "show me if i fold?", shaking nervous hands, picking
up chips as if going to bet/raise, misdeal, ... all could be nasty but also
fun.
of
weak |
|
s******g
发帖数: 15854 | 26 研究了一下，prion，misfold protein，这个东西不带核酸，基本上就是一个有机高分
子，需要非常高的温度才能破坏它的结构，明白了，不吃牛肉了。 |
|
W*******n
发帖数: 4140 | 27 我的2011年版找工简历
王利民
2014年8月16日
（现注：当时，未列出2010年在Fox Chase Cancer Center的经历；没列出2005年在
Memorial Sloan-Kettering Cancer Center的经历；没列出2006年在陕西师范大学、
2009年在江南大学任教的经历。）
Career Summary:
PhD degree on Chemistry and post-doctoral research on protein
structure in the USA; Teaching (assistance) experience in the USA and
China; Customer service experience in the USA.
Work/ Research:
Jan 2006 - : various work in the USA, China, and home.
Sept. 2003- June 2005: New York State Department of Health
Post-doctorate
... 阅读全帖 |
|
b****g
发帖数: 1204 | 28 Honor:
Highest score in the 1991' National College Entrance Exam in Hunan
Province, China
2011年版找工简历
王利民
Career Summary:
PhD degree on Chemistry and post-doctoral research on protein
structure in the USA; Teaching (assistance) experience in the USA and
China; Customer service experience in the USA.
Work/ Research:
Jan 2006 - : various work in the USA, China, and home.
Sept. 2003- June 2005: New York State Department of Health
Post-doctorate
Research projects: Crystal structure studies on superan... 阅读全帖 |
|
W*******n
发帖数: 4140 | 29 老年痴呆症赞比buydig，你何故不全文转发我的原贴呢？我全文再贴一下：
我的2011年版找工简历
王利民
2014年8月16日
（现注：当时，未列出2010年在Fox Chase Cancer Center的经历；没列出2005年在
Memorial Sloan-Kettering Cancer Center的经历；没列出2006年在陕西师范大学、
2009年在江南大学生物工程学院任教的经历。）
Career Summary:
PhD degree on Chemistry and post-doctoral research on protein
structure in the USA; Teaching (assistance) experience in the USA and
China; Customer service experience in the USA.
Work/ Research:
Jan 2006 - : various work in the USA, China, and home.
Sept. 2003- June 2005: New York... 阅读全帖 |
|
x**i
发帖数: 2627 | 30 刷屏挺好。。。
Honor:
Highest score in the 1991' National College Entrance Exam in Hunan
Province, China
2011年版找工简历
王利民
Career Summary:
PhD degree on Chemistry and post-doctoral research on protein
structure in the USA; Teaching (assistance) experience in the USA and
China; Customer service experience in the USA.
Work/ Research:
Jan 2006 - : various work in the USA, China, and home.
Sept. 2003- June 2005: New York State Department of Health
Post-doctorate
Research projects: Crystal structure studies o... 阅读全帖 |
|
b****g
发帖数: 1204 | 31 【 以下文字转载自 WaterWorld 讨论区 】
发信人: buydig (digbuy), 信区: WaterWorld
标 题: 这王利民是混的最惨的高考状元吗？
发信站: BBS 未名空间站 (Tue Aug 19 22:40:39 2014, 美东)
Honor:
Highest score in the 1991' National College Entrance Exam in Hunan
Province, China
2011年版找工简历
王利民
Career Summary:
PhD degree on Chemistry and post-doctoral research on protein
structure in the USA; Teaching (assistance) experience in the USA and
China; Customer service experience in the USA.
Work/ Research:
Jan 2006 - : various work in the USA, China, and ho... 阅读全帖 |
|
j****i
发帖数: 68152 | 32 【 以下文字转载自 WaterWorld 讨论区 】
发信人: buydig (digbuy), 信区: WaterWorld
标 题: 这王利民是混的最惨的高考状元吗？
发信站: BBS 未名空间站 (Tue Aug 19 22:40:39 2014, 美东)
Honor:
Highest score in the 1991' National College Entrance Exam in Hunan
Province, China
2011年版找工简历
王利民
Career Summary:
PhD degree on Chemistry and post-doctoral research on protein
structure in the USA; Teaching (assistance) experience in the USA and
China; Customer service experience in the USA.
Work/ Research:
Jan 2006 - : various work in the USA, China, and ho... 阅读全帖 |
|
s******y
发帖数: 28562 | 33 Wow, that is a nice discovery.
There are many genes that are controlled by temperature.
And that really depends on what do you define as "温度高" in your
experiment.
If your "high temperature" are referring to abnormally high temperature (
such as 42oC for human cell), then I would suggest you to look at signal
pathways related to misfolded protein.
，无
（只 |
|
d******t
发帖数: 65 | 34 非常谢谢你的指点。
看来你对这个领域比较了解，和我收集到的信息比较一致。
个人感觉我现在缺的就是计算机方面的背景。我想如果今后做bioinformatics或者
biomedical informatics的话，计算机方面的背景和知识非常重要，生物方面的知识因
为很多年的耳濡目染，如果某方面的知识欠缺的话，可以很快补上。所以我想加强计算
机方面的知识，掌握这个工具，然后做起事情来就会顺手的多。
因为我现在做的是Neurodegerative diseases (Huntington's Disease),这是一种
protein misfolding 引起的疾病。因为疾病的发病机制非常复杂，现在的生物研究感
觉有点小打小闹的感觉，研究某个或者几个蛋白或者基因的功能试图攻克某个疾病，个
人感觉不太可能，有点带着运气的成分。疾病的攻克需要很强大的计算机等信息技术才
能解决，因此我今后想做蛋白折叠与疾病之间的关系这个方向。
读博期间方向非常重要，不过我认为还是学习阶段。因为实验室和导师的限制，也许不
能够做自己最想做的。也许在博士毕业后，可以去合适的实验室做博后，做自己最想做
的研究和方向。... 阅读全帖 |
|
s********n
发帖数: 2939 | 35 就像楼上所说，可能是GST增加了你的蛋白的溶解度，而在pEXP5表达载体上表达的可能
是misfolded，然后被host降解了，所以没有看到inclusion body。
还是那句话，如果你需要的蛋白量不大(100-500 ug)，用leaky expression也许就够用
了，或者用GST载体表达，然后用protease切掉GST Tag。如果你需要的量比较大（>1
mg），你可能就需要做一些蛋白表达条件的优化，比如temperature, host, inducer
concentration, chaperones, tags etc.。 |
|
e****s
发帖数: 1125 | 36 你这里说的和我理解的allosteric effect有点点不一样，我对这些确实不大了解，可
能理解有偏
差。 我理解的是第3者的远程的结合引起一个蛋白构象的改变，从而影响和另一个蛋白
或者底物的结
合。另外一点，LZ可以看一下你的蛋白是单聚体还是多聚体。通常认为oligomers 相对有
allosteric effect的机率更大。
你说的这个现象肯定存在，但更像是misfolding带来的？所以不管怎么突变，一定要检
测该蛋白的主
要活性有否明显的变化，酶起码测酶活，蛋白起码检测主要功能,CD之类的也行。标记
前后也得比较活
性。
话也说回来，有一天心血来潮想了个Idea，想做两个蛋白间的结合，结果发现以前发表
的结合位点完全
错误。于是就来兴趣想确定这两个蛋白的相互结合位点，结果1年半了，还没有Promising的
结果。估计那
个大牛组里的也是逼急了，编了个故事出来。当然走了不少弯路，组里也没人做这个的。 |
|
w********r
发帖数: 1431 | 37 无语了，好多人真是擅长把简单的问题复杂化来显示自己的渊博...
你们最早是用什么Assay发现AB结合的？
那你就继续用这个Assay(不管是co-ip还是pull down还是NMR还是荧光啥高科技)，
根据已知的AB的结构做各种truncation/deletion（避免蛋白misfolding）,
继续用你的Assay检测，就知道那个部分是负责结合的了 |
|
g*****p
发帖数: 451 | 38 你说的完全是两个不同的东西
inclusion body通常是蛋白在开始fold的时候就没对，然后疏水区捣腾到一块去，这个
时候沉淀下来的东西是misfold的，这个玩意加点一般的溶剂没法溶解
蛋白沉淀有时候只是盐析或是过饱和，这种情况加点溶剂说不定还能溶解
比如给本科生做的硫酸铵分级沉淀提纯蛋白实验 |
|
C*******e
发帖数: 4348 | 39 你跟我说的才是不同的东西
我不是说加点溶剂什么的就溶解了
我是说当纯化一个蛋白的时候发现不在crude extract里在pellet的时候
不能笼统的归结为是因为“inclusion body”
有时候是纯化方法不合适
当然，蛋白一开始就没有正常折叠也会是一个可能
另外我觉得“inclusion body”的定义有待商榷
如果把E. coli里过表达的蛋白局部浓度过高的现象都叫“inclusion body”的话
那很多情况下那些蛋白根本就不是misfold的
“inclusion body通常是蛋白在开始fold的时候就没对，然后疏水区捣腾到一块去”
——这其实是一个从实验现象出发的推测
如果给你一个电镜照片
下面的这个，这是inclusion body
不过如果过表达蛋白是可溶的，然后也拿去做个电镜，说不定看起来是一样的
只是没有人这么无聊去做这样的电镜而已 |
|
g*****p
发帖数: 451 | 40 你没明白我的point的，不过无所谓了，这个我不纠缠
不过你给的电镜照片可说明不了任何问题
你给的这个照片是负染吗？我看这分辨率估计也得快几十到几百nm了吧
misfold的蛋白和局部浓度过高沉淀到一块的怎么你也得几十埃的
分辨率才好指认吧
否者那不怎么看都一样的么 |
|
C*******e
发帖数: 4348 | 41 “misfold的蛋白和局部浓度过高沉淀到一块的怎么你也得几十埃的
分辨率才好指认吧
否者那不怎么看都一样的么”
——纯好奇，有人做这个研究看有什么不同么？
我是真的很想知道
只是现在如果遇到纯化问题
就说是因为inclusion body
当然也不会专门做电镜验证是成包涵体了
毕竟那不是纯化蛋白的目的
不解决问题 |
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s********n
发帖数: 2939 | 43 18个cysteine，估计有不少的disulfide bonds，你是表达在E. coli的cytoplasm？如
果是的话，misfold的可能性很大。GST增加了fusion的可溶性，切掉以后就沉淀了。 |
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k***r
发帖数: 100 | 44 I agree with Ankyrin, looks like endosomes and lysosomes (try EEA1 and some
lysozyme markers). Some times, truncated protein after translation it
misfolds and directly sent to lysosome for degradation. |
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s******y
发帖数: 28562 | 45 呵呵呵，很多人发这些什么衰老的文章都是用来骗funding 的。
人类的衰老，我觉得本质上是个代谢问题，各种代谢废物（misfolded protein,
somatic mutant, oxidative stress, harmful matabolites, blah blah) 积累多了导
致的功能衰竭呗。
但是如果这么讲的话会骗不到funding, 所以大家都拼命的往分子机制上靠，个个都声
称他的那个pathway 是防止衰老的金钥匙。 |
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y**********n
发帖数: 478 | 46 protein folding/misfolding |
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b*****z
发帖数: 163 | 47 【 以下文字转载自 Postdoc 讨论区 】
发信人: binbinz (krystle), 信区: Postdoc
标 题: Paper Help!包子酬谢
发信站: BBS 未名空间站 (Wed Feb 29 13:46:25 2012, 美东)
Title:
High Molecular Weight Proteins and Protein Complexes: Structural Studies in
Solution
Author:
Vitali Tugarinov
DOI: 10.1002/9780470034590.emrstm1238
URL:
http://onlinelibrary.wiley.com/doi/10.1002/9780470034590.emrstm
麻烦发到
m********[email protected]
谢谢！ |
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i***0
发帖数: 160 | 48 Curr Opin Drug Discov Devel. 2009 Sep;12(5):666-81.
Small-molecule modulation of cellular chaperones to treat protein misfolding
disorders.
Sloan LA, Fillmore MC, Churcher I.
Thank you very much!!! |
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i***0
发帖数: 160 | 49 that is what everybody want for misfolded protein diseases. According to my
knowledge, there is no compounds directly boosting proteasome function. But
you should thank me that I found during my digging for the unfolded protein
response and ER stress, the Hsp90 inhibitors induce transcription of
molecular chaperones and enhance proteasome activity. It is always easier to
find an inhibitor targeting an enzyme system, but difficult for activators.
I bet you will be reach, if you find a proteasome ... 阅读全帖 |
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s******y
发帖数: 28562 | 50 能不能推荐一下 Protein misfolding 领域的review article?
my
But
protein
to
activators. |
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