C***2
发帖数: 62 | 1 Hey, I am going to generate GeneA+GeneB Knockin mouse in one locus.Furin
cleavage site plus P2A or T2A linker are chosen to link GeneA and GeneB.
This way, co-translation of GenaA and GeneB under endogenous promoter should
be achieved. I googled Furin cleavage site sequence, but i can't find out
accurate and reliable sequence ever used in vivo. So, if someone used or
know furin cleavage site, would you please send me the information and
reference? Moreover, i just read the furin cleavage, do... 阅读全帖 |
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m******5
发帖数: 1383 | 2 在做一个蛋白，用N端tag或者anti N - peptide的抗体能看到两条等量的带，一条高一
些，一条低一些，用anti C terminal的抗体就只能看到高的那条带。 因此怀疑低的带
是cleavage产物，类似pro caspase 变成caspase的过程，请教有哪些常用生物信息软
件能找到cleavage识别位点？ |
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p****p
发帖数: 3360 | 3 It seems the "cleavage" happens closer to the C-terminus.
I think alternative splicing of the mRNA is a more likely explanation of
your results. Run an RT-PCR to test it. |
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a****t
发帖数: 293 | 5 谢谢！我不知道该怎么形容这茄子有一条一条的沟，皱皮确实好像不太准确。
要不叫事业线茄子？英文名 cleavage eggplant... |
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P*****t
发帖数: 4978 | 6 Three Boobs
In our awe over the wild costumes at Comic Con 2012, we skipped over perhaps
the wildest one of all: Kaitlyn Leeb's sexy red costume and her daring
cleav -- wait a second. Does she have... three breasts?
Indeed, Leeb showed up to Comic Con sporting triple cleavage, thanks to
prosthetics. Leeb, a Toronto-based model and actress, stars as a "seductive
woman" in the upcoming "Total Recall" remake starring Colin Farrell, Kate
Beckinsale and Jessica Biel.
Schwarzenegger fans will recall t... 阅读全帖 |
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M*P
发帖数: 6456 | 7 亮点真多。三个乳房怎么搞出triple cleavage?
另外这个女的还是中国和爱尔兰的血统。外f 的后代。
perhaps
★ 发自iPhone App: ChineseWeb - 中文网站浏览器 |
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B********e
发帖数: 19317 | 8 【 以下文字转载自 Belief 讨论区 】
发信人: BlizzFlare (冰火), 信区: Belief
标 题: Christians and Cleavage (youtube)
发信站: BBS 未名空间站 (Sat Jan 2 20:29:47 2010, 美东)
重点关注第36到38秒，还有1:07到1:14。
2:38到2:39有一个笑点。 |
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t********m
发帖数: 2 | 9 I had the same problem once. No good way to improve it. Two pieces of
suggestion:
First, try different amount of thrombin, different incubation time and
different temperature.
Second, try different enzyme.
My problem was solved by adding a TEV cleavage site between the GST and my
target protein. TEV is a very sequence specific protease. My protein was
purified by glutathione column and eluted with TEV enzyme. It worked. |
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P*****0
发帖数: 436 | 10 60bases的合成的DNA，
在NaCl和MES的buffer里，想做peperidine+formate 的A-G cleavage
之前必须除盐么？
还是无所谓？ |
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b****r
发帖数: 17995 | 11 我们现在怀疑一个蛋白被caspase1 切，而且用western和一种caspase1 inhibitor证实了
现在想再用另一种方法double confirm一下。我们的想法是突变掉这个cleavage site
。不知道哪位对casepase比较有经验的，应该怎么突变最符合领域里大家的标准？或者
用什么别的方法更为大家接受/更简单？
每个有信息的回帖5个包子感谢吧 |
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l**y
发帖数: 595 | 12 以前一直放，但是觉得不加可能也没事吧，毕竟是在强酸TFA下cleavage 的 |
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S*******l
发帖数: 4637 | 13 我有个新解释。
RatG13和山甲冠状病毒本来有furin cleavage site,但是后来丢了。下面的文章能解释。
OURNAL OFVIROLOGY, June 2008, p. 6078–6083Vol. 82, No. 120022-538X/08/$08.
000 doi:10.1128/JVI.00074-08Copyright © 2008, American Society for
Microbiology.
Cleavage of Group 1 Coronavirus Spike Proteins: How Furin Cleavage Is Traded
Off against Heparan Sulfate Binding upon Cell Culture Adaptation
C. A. M. de Haan, B. J. Haijema, P. Schellen, P. Wichgers Schreur, E. te
Lintelo, H. Vennema, P. J. M. Rottier
A longstanding enigm... 阅读全帖 |
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s*******2
发帖数: 598 | 14 See below 1997 paper
Biochem J. 1997 Apr 1;323 ( Pt 1):233-7.
Actin is cleaved during constitutive apoptosis.
Brown SB, Bailey K, Savill J.
SourceDivision of Renal and Inflammatory Disease, Department of Medicine,
University Hospital, Nottingham NG7 2UH, UK.
Abstract
Proteases play an important role in the programme of cell death by apoptosis
but little is known of the substrates cleaved, particularly in constitutive
models of this type of cell death. Neutrophils spontaneously undergo
apoptosis ... 阅读全帖 |
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l******g
发帖数: 1623 | 15 感谢各位的讨论, 挺有启发, 特别是思你特提的第二点,从酶动力学上动脑筋. 我早就
该找本酶学书来看了,以前学的都忘记了.
这蛋白我已经作的很纯了, load 5 微克,跑SDS-PAGE, 只有这蛋白和它切了的fragment
. cleavage site也map出来了, 作的N-term AA sequencing. 突变cleavage site不怎
么影响cleavage, 但突变离开这site几个AA的地方,有不被cleave的.
在E.coli里面表达出来是包含体里面, 纯化很简单, refolding后也会有cleavage, pH8
.5 比较抗降解. 最近作RP HPLC, load上柱前有90%左右intact, 下来后>90%都被降解
了. RP HPLC的buffer pH 4. 有没有什么protease是在酸性下活性高的? |
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c********n
发帖数: 225 | 18 关于韩春雨NBT论文重复性的问题
【记录历史】需要记录一下：
1 沈啸老师没有原始数据，也无法重复验证NgAgo基因编辑实验
- 沈老师对于NBT论文中的数据和结论就没有发言权了
- 这样情况下，还是否应当在文章作者list上？
- 这样情况下，还是否应当在专利拥有者list上？
2. NBT文章的fig 4， fig 5 是没有人能重复出来的
- 这两个图支持 NgAgo in vivo gene editing acitivity
- 按作者贡献，是韩春雨做的实验
- 原始数据应当韩春雨拥有
3. 有没有人能够重复出来 NgAgo 的in vitro DNA nick、DSB activity？
- NBT文章的fig 1， fig 3a，b，supplementary fig 1， supplementary fig 2
- 按 文章贡献，应当是高峰做的实验
- 原始数据高峰应当拥有
- 高峰有没有自己重复过这些实验呢？
4. 有没有人能够重复出来 NgAgo 的in vivo cleavage activity？
- NBT文章的fig 3c,3d
- 按 文章贡献，应当是... 阅读全帖 |
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发帖数: 1 | 20 第一个建议真的很有用 我一定试试
A和B确实都是一次跨膜蛋白
我也非常怀疑是否是false positive
确实用的NP40 buffer 提全蛋白（7号针头抽吸）
但现在结果是A和B的全长和cleavage都作用
而且cleavage要比全长作用强很多 且cleavage在胞内部分
所以我想这样B cleavage是否就不算膜蛋白 也就不行成micelle
为了避免这种可能 所以做截段看作用位点 这样也能抛开膜蛋白特性
不管如何还是希望尽快确定是false还是true
RIPA |
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发帖数: 1 | 21 有了PRRA 基本可以排除实验室泄露可能被排除，可以转守围攻了
"There are also documented instances of the laboratory acquisition of SARS-
CoV-1 by laboratory personnel working under BSL-2 containment29,30. We must
therefore consider the possibility of a deliberate or inadvertent release of
SARS-CoV-2. In theory, it is possible that SARS-CoV-2 acquired the observed
RBD mutations site during adaptation to passage in cell culture, as has
been observed in studies with SARS-CoV5 as well as MERS-CoV31. However, the
acquisition of ... 阅读全帖 |
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d**********n
发帖数: 3634 | 22
wait, lemme see if I follow.
you are squeezing your forearm to create cleavage, like, elbow cleavage.
and it's hot.
I don't think I follow.
or maybe you were saying people use forearms under their boobs to boost
cleavage by squeezing them |
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r**r
发帖数: 171 | 23
CDC2/cycB or Erk2 or other kinases, but they do exclude some kinase like
polo,auroraA and CaMKII by in vitro kinase assay.
they do show some data in fig2A, they mutated the catalytic site of
separase and found that separase can not be cleaved. "As the active site lies
far from where cleavage occurs, this result implies that the cleavage of
separase is autocatalyzed" Not quite convincible.
If you are interested, you can go read "anaphase specific auto-cleavage of
separase" done by same |
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M****e
发帖数: 70 | 24 to
i think it requires knowledge about the structure of separase. if
intermolecular, there should be protein-protein interaction, and mutations
that inhibit the interaction should abolish the cleavage, otherwise, not. i do
not know how those genius found out the intramolecular cleavage in ribozymes,
yet i think that sequence-specific mutations help to distinguish "inter" from
"intra" activation. for "intra", the problem is to find how the cleavage
occurs, there should be significant conformation |
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l****n
发帖数: 711 | 25 那些peptide是non-specific cleavage引起的，这个很常见，能占到快20%，搜索的时
候一般都会包括这些partial cleavage的peptides。peptide的两端里有一端是non-
specific cleavage问题不大。可信不可信主要看score， 和重复出现的次数没什么关
系。 |
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s****t
发帖数: 461 | 26 30 years ago, some famous scientists faced the similar problem to yours.
Later, they solved the problem and their work got them Nobel price. Yes, I
am talking about RNA self splicing.
Unfortunately, you probably can't chemically synthesize your protein in full
-length. So it's almost impossible to get a truly clean system to demonstate
it.
Here are some suggestions:
1. Try in-vitro translation system to see if it's still the case.
2. If you can map the cleavage site, you can synthesize some pept... 阅读全帖 |
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S**********e
发帖数: 1789 | 27 给你看看我们系刚进来的助理教授
Vanderbilt University School of Medicine
Fingleton, Barbara Mary , Ph.D. Assistant Professor of Cancer Biology
Publications
Barrett, CW, Fingleton, B, Williams, A, Ning, W, Fischer, MA, Washington, MK
, Chaturvedi, R, Wilson, KT, Hiebert, SW, Williams, CS. MTGR1 is required
for tumorigenesis in the murine AOM/DSS colitis-associated carcinoma model.
Cancer Res, 71(4), 1302-12, 2011.
Koon, HB, Fingleton, B, Lee, JY, Geyer, JT, Cesarman, E, Parise, RA, Egorin,
MJ, Dezube, BJ... 阅读全帖 |
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发帖数: 1 | 31 果然是哥德巴赫猜想
作者说如果PRRA是自然进化的话，那么该动物种群的密度非常大才能有效进行自然筛选
，会是什么动物呢？
"In addition, the pangolin CoV does not carry a polybasic cleavage site
insertion. For a precursor virus to acquire the polybasic cleavage site and
mutations in the spike protein suitable for human ACE2 receptor binding, an
animal host would likely have to have a high population density – to allow
natural selection to proceed efficiently – and an ACE2 gene that is similar
to the human orthologue. Further characterization of CoVs in pan... 阅读全帖 |
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发帖数: 1 | 32 是你应该多读读书了
【 以下文字转载自 Military 讨论区 】
发信人: ntkrnl (ntkrnl), 信区: Military
标 题: 往sars里面加Furin蛋白的论文两篇
发信站: BBS 未名空间站 (Wed Feb 19 20:56:08 2020, 美东)
看来这个根本从原理上还是技术上都不难
其中一个作者是研究老鼠肝炎的
Furin cleavage of the SARS coronavirus spike glycoprotein enhances cell-cell
fusion but does not affect virion entry.
https://www.ncbi.nlm.nih.gov/pubmed/16519916
Activation of the SARS coronavirus spike protein via sequential proteolytic
cleavage at two distinct sites.
https://www.ncbi.nlm.nih.gov/pubmed/19321428 |
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g******7
发帖数: 1739 | 33 【 以下文字转载自 Fashion 讨论区 】
发信人: YourQueen (妖猴，以德服人), 信区: Fashion
标 题: 胸大挺倒霉的
发信站: BBS 未名空间站 (Wed Jul 7 15:05:43 2010, 美东)
呢穿个浅v领，都会漏cleavage.
而且适用穿的衣服少很多。。很多ruffle就穿不出好看的效果。baby doll的别人穿着清纯，你穿就是孕妇
上周末买的anthpology的top,今天穿上班，被女同事说：very cute...shows cleavage
.虽然不是恶意，但是自己一看可不是么。
可是我试穿的时候就没有觉得明显，不然也不会买。
还有前一阵买的bcbg white ruffle top。几乎也杯具了。都想转了。。还没转过东西。
不是bso，决定减肥减胸 |
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l*******i
发帖数: 153 | 34 这么说，对学生公平吗？Faculty都是这么不讲证据，碰到麻烦了，为了推卸责任而踩
自己的学生吗？
好歹去看看韩的文章，contributions中明确注明了每个人负责的实验；目前，重复不
来的那个Figure 4恰恰是韩春雨本人做的。
下面是从韩的文章copy来的，省的有些人不做功课就乱说 （genome editing即Figure
4是韩文章中最有价值和核心的结果）：
Contributions
C.H. conceived the study and designed the experiments. X.Z.S. provided
intellectual advice on the project and experimental design. C.H. performed
mammalian genome editing. F.G. performed the BLAST search and the in vitro
cleavage experiments. F.G., F.J. and Y.W. designed and constructed the
clone... 阅读全帖 |
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g****t
发帖数: 31659 | 35 来读书吧：
Saudi-Iranian Collaboration: A Forgotten Story
On January 2, 2016, the Sunni government of the Kingdom of Saudi Arabia (KSA
) executed the leading imam of the Shia community in KSA. The Shiite
government of Iran denounced this execution, as did governments throughout
the world, and avowed there would be consequences. Since that time, the
rhetoric has continued to escalate, and the world politicians and media have
talked of a possible direct war between Saudi Arabia and Iran. Almost
everyon... 阅读全帖 |
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T**7
发帖数: 264 | 36 杂志是皇家化学学会的杂志，印象因子是6。
想审的同学请发给我：
1.名字
2.email
3.expertise 关键词（5个左右）
4.发表的文章
谢谢关注，希望大家好运
题目：Redox-triggered intracellular dePEGylation based on diselenide-linked
polycations for DNA delivery
ABSTRACT: Extracellular stability to protect DNA against nucleases and
stimuli-triggered intracellular DNA release are key factors in designing non
-viral gene vectors. In this study, diselenide-linked polycation mPEG-SeSe-
PEI was developed as a new type of PEG-detachable gene vector for redox-
responsive gen... 阅读全帖 |
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d*****e
发帖数: 7368 | 37 【 以下文字转载自 Fashion 讨论区 】
发信人: YourQueen (妖猴，以德服人), 信区: Fashion
标 题: 胸大挺倒霉的
发信站: BBS 未名空间站 (Wed Jul 7 15:05:43 2010, 美东)
呢穿个浅v领，都会漏cleavage.
而且适用穿的衣服少很多。。很多ruffle就穿不出好看的效果。baby doll的别人穿着清纯，你穿就是孕妇
上周末买的anthpology的top,今天穿上班，被女同事说：very cute...shows cleavage
.虽然不是恶意，但是自己一看可不是么。
可是我试穿的时候就没有觉得明显，不然也不会买。
还有前一阵买的bcbg white ruffle top。几乎也杯具了。都想转了。。还没转过东西。
不是bso，决定减肥减胸 |
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j**4
发帖数: 10425 | 38 the cuntiness of some women just amazes me to no end
发信人: YourQueen (妖猴，以德服人), 信区: Fashion
标 题: 胸大挺倒霉的
发信站: BBS 未名空间站 (Wed Jul 7 15:05:43 2010, 美东)
呢穿个浅v领，都会漏cleavage.
而且适用穿的衣服少很多。。很多ruffle就穿不出好看的效果。baby doll的别人穿着
清纯，你穿就是孕妇
上周末买的anthpology的top,今天穿上班，被女同事说：very cute...shows cleavage
.虽然不是恶意，但是自己一看可不是么。
可是我试穿的时候就没有觉得明显，不然也不会买。
还有前一阵买的bcbg white ruffle top。几乎也杯具了。都想转了。。还没转过东西。
不是bso，决定减肥减胸 |
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Y*******n
发帖数: 4646 | 39 呢穿个浅v领，都会漏cleavage.
而且适用穿的衣服少很多。。很多ruffle就穿不出好看的效果。baby doll的别人穿着清纯，你穿就是孕妇
上周末买的anthpology的top,今天穿上班，被女同事说：very cute...shows cleavage
.虽然不是恶意，但是自己一看可不是么。
可是我试穿的时候就没有觉得明显，不然也不会买。
还有前一阵买的bcbg white ruffle top。几乎也杯具了。都想转了。。还没转过东西。
不是bso，决定减肥减胸 |
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A********r
发帖数: 2640 | 40 V领就是给有cleavage的美女穿的~
猴猴你还说不是BSO，发指！！！
着清纯，你穿就
是孕妇
cleavage
西。 |
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b*****e
发帖数: 14299 | 41 ☆─────────────────────────────────────☆
Vinic (Vinic) 于 (Fri May 28 10:40:28
2010, 美东) 提到:
迎合节日的喜庆气氛,限时放送美女啦~~~~~~girls'only event at my apt ;-))
过完节了，删了照片；－）））
☆─────────────────────────────────────☆
ifdef (ifdef) 于 (Fri May 28 10:41:53
2010, 美东) 提到:
ohh，my！
☆─────────────────────────────────────☆
bethoven (千金散尽还复来) 于 (Fri May 28
10:43:18 2010, 美东) 提到:
赞，美女拍的很好啊
165.89.]
☆─────────────────────────────────────☆
smugmug (摄影—记录生活中的点点滴滴) 于 (Fri May
28 10:48:19 2010, 美东) 提到:
ero... 阅读全帖 |
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l***a
发帖数: 12410 | 42 【 以下文字转载自 Fashion 讨论区 】
发信人: YourQueen (妖猴，以德服人), 信区: Fashion
标 题: 胸大挺倒霉的
发信站: BBS 未名空间站 (Wed Jul 7 15:05:43 2010, 美东)
呢穿个浅v领，都会漏cleavage.
而且适用穿的衣服少很多。。很多ruffle就穿不出好看的效果。baby doll的别人穿着清纯，你穿就是孕妇
上周末买的anthpology的top,今天穿上班，被女同事说：very cute...shows cleavage
.虽然不是恶意，但是自己一看可不是么。
可是我试穿的时候就没有觉得明显，不然也不会买。
还有前一阵买的bcbg white ruffle top。几乎也杯具了。都想转了。。还没转过东西。
不是bso，决定减肥减胸 |
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p****i
发帖数: 5597 | 43
其实我也没明白阿婆嘛意思
cleavage为啥得用前臂啊
cleavage用push up bra就有了。。。。 |
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s*******d
发帖数: 17566 | 44 【 以下文字转载自 Fashion 讨论区 】
发信人: YourQueen (妖猴，以德服人), 信区: Fashion
标 题: 胸大挺倒霉的
发信站: BBS 未名空间站 (Wed Jul 7 15:05:43 2010, 美东)
呢穿个浅v领，都会漏cleavage.
而且适用穿的衣服少很多。。很多ruffle就穿不出好看的效果。baby doll的别人穿着清纯，你穿就是孕妇
上周末买的anthpology的top,今天穿上班，被女同事说：very cute...shows cleavage
.虽然不是恶意，但是自己一看可不是么。
可是我试穿的时候就没有觉得明显，不然也不会买。
还有前一阵买的bcbg white ruffle top。几乎也杯具了。都想转了。。还没转过东西。
不是bso，决定减肥减胸 |
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w***y
发帖数: 1313 | 45 cleavage是啥？
着清纯，你穿就是孕妇
cleavage
西。 |
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s****r
发帖数: 2386 | 46 nnd, remember the other day we talked about 毛叔peeking on the cleavage when
taking picture with 大王, a few nights ago I had a dream where I was
walking downhill at some hiking place, I kept peeking into girls' cleavages
while climbing down... |
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g*****p
发帖数: 451 | 47 It will, for sure, if there is an accessible cleavage site in itself.
Auto-cleavage is quite common in the maturation process of zymogen,
particularly during viral protease maturation. |
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g*****p
发帖数: 451 | 48 trans-cleavage does bite; but sometimes, even cis-cleavage is also possilbe |
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S**********e
发帖数: 1789 | 49 看了这个教授的主页，竟然公布了自己review的文章，题目，杂志名称都有，是不是违反职业道德呢？
Editorial Activities
Journal Peer Reviewer
2009 International Journal of Cancer. (Article) E-cadgerin
transcriptional down-regulation by epigenetic and microRNA-200 family
alterations is related to mesenchymal and drug-resistant phenotypes in human
breast cancer cells, IJC-09-1835.
2009 Molecular Cancer Therapeutics. (Article) Role of reactive oxygen
species in the ability of H-Ras to enhance cell death induced by histone
deacetylase inhibit... 阅读全帖 |
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