D*****y
发帖数: 5788 | 1 ☆─────────────────────────────────────☆
jym2307 (罪魁) 于 (Tue Dec 7 19:14:10 2010, 美东) 提到:
从哪里说起都可以
只有一条，最好代表当今进化论的权威
否则不要天天浪费大家的时间跟你一起打酱油
基督版不是给你进化论妄想打酱油的地方
☆─────────────────────────────────────☆
jym2307 (罪魁) 于 (Tue Dec 7 19:16:08 2010, 美东) 提到:
给你三天时间放狗去找
够了吧？
☆─────────────────────────────────────☆
atep (嘚嘚) 于 (Tue Dec 7 23:28:47 2010, 美东) 提到:
请最虔诚的罪魁来说说基督教-三日为限
从哪里说都可以
该不该蒙头 该不该抹油 该不该搞基 该不该吃转基因食物
只有一条 要代表当今基督教的权威答案 好让这些问题有统一答案
给你三天时间 放狗去搜吧
☆─────────────────────────────────... 阅读全帖 |
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t*******d
发帖数: 2570 | 2 Orig Life Evol Biosph. 2008 Feb;38(1):57-74. Epub 2007 Nov 16.
Lipid-assisted synthesis of RNA-like polymers from mononucleotides.
Rajamani S, Vlassov A, Benner S, Coombs A, Olasagasti F, Deamer D.
Source
Department of Chemistry and Biochemistry, University of California, Santa
Cruz, CA 95064, USA.
Abstract
A fundamental problem in research on the origin of life is the process by
which polymers capable of catalysis and replication were produced on the
early Earth. Here we show that RNA-like poly... 阅读全帖 |
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t*******d
发帖数: 2570 | 3 Orig Life Evol Biosph. 2008 Feb;38(1):57-74. Epub 2007 Nov 16.
Lipid-assisted synthesis of RNA-like polymers from mononucleotides.
Rajamani S, Vlassov A, Benner S, Coombs A, Olasagasti F, Deamer D.
Source
Department of Chemistry and Biochemistry, University of California, Santa
Cruz, CA 95064, USA.
Abstract
A fundamental problem in research on the origin of life is the process by
which polymers capable of catalysis and replication were produced on the
early Earth. Here we show that RNA-like poly... 阅读全帖 |
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O**********0
发帖数: 61 | 4 ☆─────────────────────────────────────☆
BHistory (往者的影子) 于 (Fri Sep 9 00:20:11 2011, 美东) 提到:
俺曾认为生命是进化起源的，后来越找越发现进化论像是信仰而非科学结论。可是每有
飞机网友觉着俺如此看待进化论是反科学了。
所以俺本着科学的精神，问个问题，求各位飞机进化论牛人给个科学的答案。
大家知道达尔文进化论是以遗传机制为基础的，第一个有遗传机制的生命怎么来的，当
时根本没有提及。
后人发现这一问题后，就又提出了化学进化论，就是无机分子在某种机制下，自动组合
出了第一个有遗传的生命。此后达尔文进化论似乎可以take care（其实未必，今后再
讨论）。
今天我们先看比第一个有遗传生命的出现简单万亿倍的问题，第一个蛋白质如何化学进化出
现？
先亮亮俺一个愚昧基督徒的愚见：
已知最简单的具有遗传机制的单细胞生命的复杂度已经远远超出人们最初的想象，大概
有400种蛋白质几十万个组成。其中较小的蛋白质大概100个氨基酸，如果在随机热运动
中组成，因为自然界对称的氨基酸不少于40种（20多种氨基酸及... 阅读全帖 |
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O**********0
发帖数: 61 | 5 ☆─────────────────────────────────────☆
BHistory (往者的影子) 于 (Fri Sep 9 15:00:23 2011, 美东) 提到:
本帖是热运动组装第一个蛋白质的副贴。原帖：http://www.mitbbs.com/article_t1/TrustInJesus/676133_0_1.html
因为靠热运动组装第一个蛋白质是困难的，所以有观点认为第一个蛋白质是根据某个自
然产生的遗传基因密码(DNA/RNA)组装出来的。这便是热运动组装第一个遗传密码/基因
的理论。
在俺看来，这是先有蛋还是先有鸡的问题。蛋白质好比蛋，基因好比下蛋的鸡。如果自
然界糊出一个蛋很困难，那糊出一个下那个蛋的鸡就更困难。
事实上，每三个DNA碱基对决定一个氨基酸，加上基因的头码三个，尾码三个，对应一
个100个氨基酸长链的遗传密码（306个碱基对），其自然产生的困难程度和自然排列该
特定氨基酸长链的困难程度基本一样（遗传密码更难一点）。且遗传密码翻译为第一个
蛋白质还可能需要翻译和连接氨基酸的机制（类似一些细胞器的功能）。所以自然产生
第一个... 阅读全帖 |
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c***u
发帖数: 3888 | 6 有问题啊， 毛叔给你检查下prostate和seminal vesicle |
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s******y
发帖数: 28562 | 7
Search the amino acid sequence on data base for signal peptide.
MOst signal peptides have been characterized.
I think, roughly, but not in detail: digest the whole cell or the membrane
vesicles make from supersonic, without using any detergent. If a pepetide is
outside membrane, it will be digested. If it's inside, it will not.
GFlycosylation should use similar mechanism, but instead of protease,
glycosylase is used to labled the outside peptide, then a mess spec can be
used to determined the l |
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v*****n
发帖数: 133 | 8 SNAREs can be grouped into t-SNARE (target) and v-SNARE (vesicle), or Q-SNARE
and R-SNARE based on sequence. 4 families have been found: syntaxin, SNAP-25-1,
SNAP-25-2 (Q type), and synaptobrevin (R type).
SNARE complex during membrane fusion is actually a 4 helical bundle, 2 from each
membrane location in the ratio of 3Q : 1R.
Besides this, the subunit c of the V0 sector of V-type ATPase, also called
proteolipid, is also shown to facilitate homotypic fusion in yeast, which was
published in Nat |
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m*p
发帖数: 226 | 9 Embryonic hearts were co-transfected by RFP and GFP plasmids. Only 20% were
transfected. Boss argued that
all RFP and GFP should co-localized and I said it is impossible, only when
RFP and GFP plasmids are in the
same lipid vesicle and fused with the same cell, then they will colocalize.
She said she was one thousand
percent sure. I can not say more, as this might harm the relationship.
However, I would like to know the truth about this co-transfection. Thanks
for your imput. |
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w*****y
发帖数: 35 | 10 完全不一样，autophagy vesicle里面有很多electron dense的黑黑的东西 |
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t*x
发帖数: 1065 | 11 有个别细胞没处理前确实也有vesicle
不过那是极少数的细胞，而且数目也并不多，在我看来应该是正常情况下的autophagy
不过，处理后的细胞很容易看出区别
几乎每个细胞里都有，而且随着时间的增加，数目越来越多，体积也越来越大 |
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s******y
发帖数: 28562 | 12 我用的是embryonic fibroblast, 和3T3差不多。
一般都能做到20%。 再高就没有办法了。
基本上，你要确认两件事：
1。你提的DNA足够纯。如果不放心的话，可以用ethanol/NaAc 沉淀法
纯化一次
2。细胞的状态。 必须在60~80% confluence 之间。
如果都不行的话，还有一个不是办法的办法，
那就是把细胞trpsinized, wash, resuspend, 然后在细胞贴壁之前
把转染混合物加进去。因为细胞在spread and adhere 的时候会吸进很多
vesicle.相当于就是强行把DNA弄进去了。不过这个方法会弄死很多细胞。
谢。 |
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s******y
发帖数: 28562 | 13 It's better you post the photo on the board.
Most likely the things you saw in the Hela cells are some cellular vesicles
or mitochondria |
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w**********s
发帖数: 72 | 14 Flow cytometry as a novel tool to evaluate and separate vesicles using
characteristic scatter signatures.
J Phys Chem B. 2010 Aug 12;114(31):10010-6.
Thanks you very much! |
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y******8
发帖数: 1764 | 15 One sample from google. Good luck!
Dear PLoS Biology editor:
I would like to submit a presubmission inquiry regarding suitability of our
manuscript, "Subversion of cellular autophagy pathway by RNA viruses" by
William T. Jackson, Thomas H. Giddings Jr., Sara Mulinyawe, Ron Kopito and
Karla Kirkegaard for publication in PLoS Biology. Cellular autophagy is a
field that is just entering a period of rapid discovery, because the
mammalian homologs for the many yeast genes that affect autophagy have v... 阅读全帖 |
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b****n
发帖数: 311 | 16 呵呵，看看薛红卫，去掉了20几篇plant cell以下的。
1: Wu GZ, Xue HW. Arabidopsis β-ketoacyl-[acyl carrier protein] synthase i
is
crucial for fatty acid synthesis and plays a role in chloroplast division
and
embryo development. Plant Cell. 2010 Nov;22(11):3726-44. Epub 2010 Nov 16.
PubMed
PMID: 21081696; PubMed Central PMCID: PMC3015132.
2: Dai C, Xue HW. Rice early flowering1, a CKI, phosphorylates DELLA protein
SLR1
to negatively regulate gibberellin signalling. EMBO J. 2010 Jun 2;29(11):
1916-27.
Epub 2010 Apr 16... 阅读全帖 |
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c********r
发帖数: 1125 | 17
不止一篇pnas,一篇cell metabolism。。。
Zhou KM; Dong YM; Ge Q ... Xu T, PKA activation bypasses the requirement for
UNC-31 in the docking of dense core vesicles from C.elegans neurons, Neuron
, 2007, 56: 657-669.
Bai L, Wang Y, Fan JM... Xu T. Dissecting multiple steps of GLUT4
trafficking and identifying the sites of insulin action. Cell Metabolism.
2007, 5:47-57.
Kang LJ, He ZX, Xu PY, Fan J, Betz A, Brose N, Xu T. Munc13-1 is required
for the sustained release of insulin from pancreatic beta cells. C... 阅读全帖 |
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e**r
发帖数: 1144 | 18 是在vesicle里的
张辰宇去年一篇mol cell
Secreted monocytic miR-150 enhances targeted endothelial cell migration
不仅可以保护mirna而且可以 定向运送到特定细胞
感觉这个领域前景广阔啊 |
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e**r
发帖数: 1144 | 19 是在vesicle里的
张辰宇去年一篇mol cell
Secreted monocytic miR-150 enhances targeted endothelial cell migration
不仅可以保护mirna而且可以 定向运送到特定细胞
感觉这个领域前景广阔啊 |
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w**r
发帖数: 134 | 20 I like this prediction.
http://wavefunction.fieldofscience.com/2011/09/2011-nobel-prize
MEDICINE:
1. Nuclear receptors (Easy)
Pros: The importance of these proteins is unquestioned. Most predictors seem
to converge on the names of Chambon/Jensen/Evans.
2. Chaperones: (Easy)
Arthur Horwich and Franz-Ulrich Hartl just won this year's Lasker Award for
their discovery of chaperones. Their names have been high on the list for
some time now.
Pros: Clearly important. Chaperones are not only important f... 阅读全帖 |
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b******s
发帖数: 1089 | 21 很多co-localization只是同是定位于胞内的compartments，并不能说明两个蛋白相互
接触和作用。
比如一个vesicle上可以有很多蛋白，但他们不一定要直接接触。
要直接证明，还是需要其他手段，live imaging可以用， FRET或者BIFC
生化的可以做pull down或者co-ip |
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l**********1
发帖数: 5204 | 22 //www.ncbi.nlm.nih.gov/pubmed/21050804
浮黴菌門
Isosphaera屬
//hamap.expasy.org/proteomes/ISOPI.html
Species: Isosphaera pallida (strain ATCC 43644 / DSM 9630 / IS1B)
Species code: ISOPI
Taxonomy: Bacteria; Planctomycetes; Planctomycetia; Planctomycetales; Planctomycetaceae; Isosphaera
(TaxID: 575540) [NEWT/ NCBI]
Description: Isosphaera pallida strain IS1B was isolated from a hot spring in Kah-nee-tah, Oregon, USA.
It is the only budding bacterium known to glide and the only heterotrophic bacteriu... 阅读全帖 |
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l**********1
发帖数: 5204 | 23 Please refer:
scanned Bessel beams in conjunction
with structured illumination and/or two-photon excitation to create thinner
light sheets (<0.5 μm) better suited to three-dimensional (3D) subcellular
imaging
PubMed link:
//www.ncbi.nlm.nih.gov/pubmed/21378978
Planchon TA, Gao L, et al.
Rapid three-dimensional isotropic imaging of living cells using Bessel beam
plane illumination.
Nat Methods. 2011 May;8(5):417-
>Abstract
>A key challenge when imaging living cells is how to noninvasively extract... 阅读全帖 |
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i***l
发帖数: 1656 | 24 分crude membrane and cytosolic fraction, how crude? sure 100% seperated by
centrifugation?
用cofocal看却看到在 everywhere, marker? plasma- membrane and introcellular
vesicle markers? |
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b***n
发帖数: 36 | 25 请问版上有玩ＧＰＣＲ尤其是dopamine receptor的兄弟姐妹不
我现在在做DR internalization, 试了好几种办法，
不管是Membrane biotinylation,
或者是用Flag M2在没有fix之前去label外膜上的receptor
或者是用Radio ligand来做，
都不是很成功，最近试了一下用Sucrose gradient (35%)来分heavy membrane 和light
vesicle, 应该是有看到了一些internalization.
不过最大的问题是用ＦＬＡＧ Ｍ２做完ＷＢ后看到的并不是一条单一的带，而是一片
的smear,
应该是某些地方没做好，所以Ｄ２Ｒ都聚集了
用的是Biorad的ＳＤＳ loading dye 配方直接溶解膜
或者也试过ＲＩＰＡ和ＴＢＳ＋１０％甘油＋１％triton
效果都不是太好
但是看了不少ｐａｐｅｒ貌似人家都是做得出来了
这个事情搞了大半年，一直没能搞定，老板又特别楞，从来不给些建设性的意见，每隔
一段时间都要威胁一下做不出来就开除，已经被威胁了十数次，真是有点窘境穷途了,,
,,还能不能接着... 阅读全帖 |
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k******0
发帖数: 1073 | 26 Odds Against Winning the 2012 Nobel Prize in Chemistry
Nuclear Hormone Signaling, Chambon/Evans/Jensen, 6-1
Bioinorganic Chemistry, Gray/Lippard/Holm/–, 7-1
Spectroscopy & Application of Lasers, Zare/Moerner/+, 8-1
Techniques in DNA Synthesis, Caruthers/Hood/+, 13-1
The Field (everything not listed), 14-1
Electrochemistry/Electron Transfer, Bard/Hush/Gray/–, 19-1
Biological Membrane Vesicles, Rothman/Schekman/+, 19-1
Instrumentation/Techniques in Genomics, Venter/+, 24-1
Molecular Studies of Gen... 阅读全帖 |
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n**********7
发帖数: 29 | 28 大家好。
我最近在用flow检测一种膜蛋白的表达。对于这种蛋白，现有的用于flow的抗体都是
recognize c－terminal的，所以只能做intracellular flow。我的基本步骤是分离细
胞后，加Fc block，fix，permeablize，0.5% BSA block buffer,然后分别染一抗和二
抗，只有single color。
现在遇到的问题是，这种蛋白在treatment之前，不仅在细胞表面有表达，也存在于
intracellular vesicles里，所以不知道permeablize之后会不会检测到的量不仅仅是
表面的表达量。具体应该怎么做才能够保证我仅仅检测到cell surface的量呢？或者除
了flow之外还有其他的方法更适合解决这个问题？
多谢了！
做这个实验有两个多月了，反复试结果都不好，真着急！！ |
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c********n
发帖数: 67 | 29 Inside-Out Red Cell Membrane Vesicles: Preparation and Puirification
PMID: 5418644
Thanks very much for you kindly help.
h***[email protected] |
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l**********1
发帖数: 5204 | 30 >Südhof是大牛，每个做突触递质释放的都应该知道这个人
Bingo
The Nobel Prize in Physiology or Medicine 2013 was awarded jointly to James
E. Rothman, Randy W. Schekman and Thomas C. Südhof "for their discoveries
of machinery regulating vesicle traffic, a major transport system in our
cells".
http://www.nobelprize.org/nobel_prizes/medicine/laureates/2013/ |
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s*******7
发帖数: 399 | 31 按照今年的发奖主题和工作类似性，阿戈个人认为，和Rothman，Schekman 去 share
的应该是Camilli。而Südhof则可以在未来和其他人去组合（比如更接近Neuroscience
的主题）。
当然，有的人拿奖是必然的，即使现在没有，以后也会迟早轮到。
除了今天的Südhof，跟 Neuroscience 沾边的阿戈觉得以后还会有 Julius 和
Camilli；当然，一定少不了Deisseroth。
在 soccer007 (阿戈) 的大作中提到:
Bingo
The Nobel Prize in Physiology or Medicine 2013 was awarded jointly to James
E. Rothman, Randy W. Schekman and Thomas C. Südhof "for their discoveries
of machinery regulating vesicle traffic, a major transport system in
ourcells".
http://www.nobelprize.org... 阅读全帖 |
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l******u
发帖数: 936 | 32 James E. Rothman, Randy W. Schekman
and Thomas C. Südhof
for their discoveries of machinery regulating vesicle traffic,
a major transport system in our cells |
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l******u
发帖数: 936 | 33 现在肿瘤患者/肿瘤动物模型血液中的exosome研究也很火热，这和他们的vesicle 研究
相关， 比如一些 RAB 蛋白, 在肿瘤和神经的生物学功能都非常有趣。 |
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b******s
发帖数: 1089 | 34 看今年Nobel prize有感。植物领域现在一统天下的还是characterize不同基因的功能
。在细胞学层次上，还是一团糊。很多植物领域的cellular biology的结果都不太可靠
，不少做植物细胞学的估计也有同感。
植物细胞学是比较难做，没有可靠的单细胞系统，组织自发荧光很强，imaging不好做
。而且大部分成熟的植物细胞基本被vacuole占据，没有给vesicle trafficking足够的
观测空间。 |
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w***i
发帖数: 175 | 35 更加狗血的是，还真有人把前女朋友介绍给同事去接盘。广大的接盘男们信心大振。
大侠牛B, 大学未毕业就发了三篇paper,其中包括science
UCLA alumnus Randy Schekman, a professor of molecular and cell biology at UC
Berkeley, has won the 2013 Nobel Prize in physiology or medicine for his
role in figuring out how proteins are secreted and transported in human
cells.
He shares the prize with James E. Rothman of Yale University and Thomas C. S
üdhof of Stanford University for solving the mystery of how the cell
organizes its transport system.
Schekman became the... 阅读全帖 |
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L*********y
发帖数: 322 | 36 I have some comments on this year's three nobel prize laureates in
physiology and medicine. As compared to Schekman and Rothman, Sudhof made
far less contributions to membrane traffic. It is Randy Schekman's
pioneering yeast genetic screen that led to the first identification of a
number of vesicle secretion mutants. Remarkably, these genes turned out to
be highly conserved between yeast and higher organisms including mammals.
James Rothman did very fine biochemical and in-vitro cell re-constitu... 阅读全帖 |
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s******a
发帖数: 472 | 37 肝细胞细胞质有很强的自发荧光，主要来自lipid droplets或者vesicles。如果做
imaging的话，大家都有什么trick可以减少自发荧光。
我的感觉是在体外培养时间越长自发荧光也增强。 |
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D***a
发帖数: 516 | 38 会不会是GFP因为没有binding partner， 所以最后沦落到endosome like vesicle里了
？有功能的蛋白会去它该去的地方？ |
|
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s******y
发帖数: 28562 | 40 不是这个意思啊，我才发现你和我讲的不是一回事。
我说的是NIH 不乐意资助一般的细胞生物学了，那些专门研究focal adhesion,
vesicle motility,cell motility 的人很多都拿不到资金了。
后来我也说了，细胞生物的一个出路就是细胞分化工程。
你说的这种其实就是细胞分化工程。人工材料到底是不是完全和体内的一样无所谓，只
要能把细胞诱导到想诱导的样子就行了。这种情况当然是可以拿到钱的，但是你说的这
种研究，在本质上来说，其实并不管里面的机理是什么回事。相关的人虽然也做做机理
来拓宽以后的manipulation 的思路，但是“机理”并不是这种研究项目的deliverable
items. 所以NIH 不资助那些专门搞细胞生物学的人的原因，和你说的这种研究并没有
直接关系。 |
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n**e
发帖数: 2026 | 41 【 以下文字转载自 WaterWorld 讨论区 】
发信人: nile (nile), 信区: WaterWorld
标 题: 美国科学促进会《科学》大旗下的转基因谎言
发信站: BBS 未名空间站 (Fri Oct 30 10:18:45 2015, 美东)
今年初，美国科学促进会（AAAS）和皮尤研究中心（Pew Research Center）联合公布
的一项调查显示，88％的科学家认为转基因食品可安全食用，持这一看法的公众只有37
％。报告认为差距如此之大的一个可能原因是，三分之二的公众认为科学家对转基因食
品的健康后果没有清晰认识。这就是真正的原因吗？
根据报告全文，皮尤中心给19，984名科学家发出了调查问卷，3748人（18.8%）完成了
问卷调查。其中生物医学学科背景的科学家占50%。报告没有说明50%的生物医学背景是
否能够代表科学家总体的学科构成。更大问题是在这50%的科学家人群中，有多少人的
工作是与转基因作物的研究有关，以什么样的方式相关。报告没有提供任何信息。
由于这些关键信息缺失，88%的科学家相信转基因安全的说法毫无意义。在一个没有代
表性并且无法排除... 阅读全帖 |
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发帖数: 1 | 42 Methods Mol Biol. 2016;1434:153-67. doi: 10.1007/978-1-4939-3652-6_11.
Protocols for Exosome Isolation and RNA Profiling.
Marqués-García F1,2, Isidoro-García M3,4,5.
Author information
Abstract
Exosomes are small extracellular vesicles of multivesicular bodies derived
from the cell endosome. Molecules have a characteristic composition both in
the membrane and in the carried load. These particles are located in the
culture medium of cells in vitro and in vivo body fluids, being synthesized
by alm... 阅读全帖 |
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发帖数: 1 | 43 Collecting plasma exosomes/extracellular vesicles, check exosomal RNAs (
mainly microRNAs) as well as proteins (oncogenic proteins involved in tumor
metastasis). |
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发帖数: 1 | 44 你好，本人有神经生物学背景，目前研究方向为星形胶质细胞extracellular vesicle
对神经元树突棘可塑性的调控。如有合适的文章，请楼主考虑！楼主好人，谢谢！ |
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发帖数: 1 | 45
Trying to search with "extracellular vesicles/exosomes". |
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发帖数: 1 | 46 paper thanks!
Nat Immunol. 2002 Dec;3(12):1156-62. Epub 2002 Nov 11.
Indirect activation of naïve CD4+ T cells by dendritic cell-derived
exosomes.
Théry C1, Duban L, Segura E, Véron P, Lantz O, Amigorena S.
Author information
Abstract
Dendritic cells (DCs) secrete vesicles of endosomal origin, called exosomes,
that bear major histocompatibility complex (MHC) and T cell costimulatory
molecules. Here, we found that injection of antigen- or peptide-bearing
exosomes induced antigen-specific na&... 阅读全帖 |
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C****c
发帖数: 1964 | 47 Can someone share these three papers with me? Thank you very much.
1. Nature Reviews Gastroenterology and Hepatology 10, 656-665 (November 2013
) | doi:10.1038/nrgastro.2013.183
NAFLD, NASH and liver cancer
2. Lipid droplets and liver disease: from basic biology to clinical
implications
Nature Reviews Gastroenterology & Hepatology 14, 343–355 (2017) doi:10.1038
/nrgastro.2017.32
3. Extracellular vesicles in liver disease and potential as biomarkers and
therapeutic targets
Nature Reviews Gastroen... 阅读全帖 |
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l*****7
发帖数: 2844 | 48 Could you kindly get this paper for me? Thanks a lot!
http://www.rsc.org/publishing/journals/JM/article.asp?doi=b818006g
J. Mater. Chem., 2009, 19, 2037 - 2042, DOI: 10.1039/b818006g
Ionic self-assembled solid-like vesicles and their temperature-induced
transformation
Bo Jing, Xiao Chen, Yurong Zhao, Xudong Wang, Fumin Ma and Xiu Yue
My email is l*****[email protected]
Thank you very much in advance! |
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d********y
发帖数: 176 | 49 what kind of membrane system are you using? vesicle? supported bilayer? |
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w********h
发帖数: 12367 | 50 Paul Janmey没当上呢，Dennis Discher倒先用这个citation上了。。。
后者应该加上polymer self-assembled vesicles. |
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