r*********n 发帖数: 13992 | 1 Mouse colon tissue,做了很多次了,有时候有信号,有时候没有,快被折磨疯掉了,
TNND,俺还用TSA boost呢......
看人家说的这个抗体(cell signaling)的)挺好用的阿,我做的这个咋就这么不稳定
呢。试了其他的apoptosis marker的抗体,例如Cleaved-PARP,也是没有什么信号。但
是我做别的抗体就没什么问题,结果还不错,就是不知道这个的问题到底出在哪里,烦
死人阿。
TUNEL做完了,老板非要其他一个marker作比较,唉,童鞋们你们谁做过这个?能不能
说一下关键步骤在哪里啊? |
|
i*********0 发帖数: 915 | 2 一个小问题,这个 Cleaved caspase-3 是看p53 independent apoptosis么?
我记得在一片paper上看过,现在忘记了. |
|
k******n 发帖数: 13 | 3 SiO2 has 7 types of crystals and one glassy type. If it is from single crystal
or glass, it is transparent, if from multi-crystal or from a lot of amorphous
particles, must be opaque due to the scattering.
Why not cleave and/or polish the end of your matrix with film (if it is not
too hard), and observe from the cross-section by SEM? It will be more precise
and direct than optical measurement based on te reflection or transmission
equation. |
|
P*****e 发帖数: 25 | 4 今天有空去查了一下CRISPR专利的历史,UCB的专利已经有claim通过了审查(allow)
,只是还没有issue(时间问题)。我在后面列出来了其中一个通过了的claim,同时把
张锋issue了的专利里面的一个claim也列了出来。大家看能避开不?张锋后来的Cpf1看
着能避开UCB的Cas9。
有意思的是发现最早的一篇CRISPR相关专利申请其实是Luciano A. Marraffini(张锋
合作者)在Northwestern University时申请的,priority date Sep 23, 2008,早了
好几年。看看claim,只有interference,还没进化到切割,可惜一个claim也没批,居
然华丽丽的被一堆112给毙了,也就是审查员觉得那专利写的不清不楚,业内普通人士
看着不可能做出claim的结果,连101,102,103都没用上就挂了。西北也是早早的
abandon了,当年连PCT都没申请,可见并不重视。张锋专利里也没有Marraffini,虽然
science文章有他,我猜可能也是他只做真核interference方面的工作吧,张跟他合作
并进... 阅读全帖 |
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r****1 发帖数: 2299 | 5 He had been a seemingly all-American, clean-cut guy: No criminal record.
Engaged to be married. A job teaching English overseas. In letters to the
judge, loved ones described the 29-year-old Midwesterner as honest and
caring — a good citizen. His fiancee called him "Mr. Patriot."
Such descriptions make the one that culminated in the courtroom all the more
baffling: Glenn Shriver was also a spy recruit for China. He took $70,000
from individuals he knew to be Chinese intelligence officers to try ... 阅读全帖 |
|
发帖数: 1 | 6 Argonaute (Ago) proteins are complexed with small DNA or RNA guides and
cleave nucleic 24
acid targets whose sequences are complementary with the guides1. Agos in
eukaryotes bind 25
to small interfering RNAs (siRNAs) or microRNAs (miRNAs) to suppress gene
expression by 26
degrading mRNAs in a targeted manner, a regulatory process known as RNA
interference 27
(RNAi). Agos in eubacteria and archaea are complexed with DNA or RNA guides
and function 28
as defense systems against foreign mobile genet... 阅读全帖 |
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l****z 发帖数: 29846 | 7 By JUSTIN SCHECK and IANTHE JEANNE DUGAN
BLUE LAKE, Calif.—Malodorous brown smoke from a power plant enveloped this
logging town on April 29, 2010, and several hundred residents fled until it
passed.
Six months later, the plant got $5.4 million from a federal program to
promote environmentally preferable alternatives to fossil fuel.
The plant, Blue Lake Power LLC, burns biomass, which is organic material
that can range from construction debris and wood chips to cornstalks and
animal waste. It is... 阅读全帖 |
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B********e 发帖数: 19317 | 8 【 以下文字转载自 WaterWorld 讨论区 】
发信人: ruomu (ruomu), 信区: WaterWorld
标 题: 一个学长天天玩dota 结果GRE接近满分 让你见识下。。。
发信站: BBS 未名空间站 (Fri Apr 1 10:58:57 2011, 美东)
总的来说,DoTA是一个文化包容性很强的游戏,而且DOTA中文版翻译的还是非常非常好
的~~~
(既然这么多人看。。lz顺征人交流魔兽对战,是1v1对战喔)
后文会出现的一些常用缩写:
War3:warcraft 3 魔兽争霸3
RA:red alert 红色警戒系列
SC: star craft 星际争霸
CoD:call of duty 使命召唤
还有一些游戏的名称在文中会有注释。
好了,正文:
关于Dota和一些游戏中的GRE词汇
A字头
Abaddon :DOTA中地狱领主的名字。本意为地狱,恶魔。是圣经里的亚玻伦 (地狱的
使者)。
abomination:war3中的憎恶。abominate痛恨,憎恶。
aegis:immortal aeg... 阅读全帖 |
|
y*******n 发帖数: 10103 | 9 【 以下文字转载自 Diablo3 俱乐部 】
发信人: yeemartin (猪头), 信区: Diablo3
标 题: 简短19级野蛮人感想
发信站: BBS 未名空间站 (Wed May 16 16:01:04 2012, 美东)
随便写写,d2当时也没玩很多。基本是单机。现在印象也不深了,所以就不对比了。
初期感觉这次野蛮人很强大,AOE攻击技能很多,伤害输出相当可观,感觉不算只是肉
盾角色了。几个技能的回血神符也很好用。往往好像要不行了,一下子就回血了。比如
rend和revenge这2个技能的神符。
一般是起招是ground stomp或leap,这样就有15的fury,整晕周围敌人或增加防御4秒
。我推荐ground stomp,有个神符可以把敌人拉拢,这样好用rend这种AOE。震晕这4秒
时间基本无敌,趁机cleave一下(扇面120%武器伤害),再得8点fury。然后就可以放
rend了。rend是AOE 220%武器伤害,加blood thirst神符可得转化9%伤害为血,这个很
爽,持续rend基本就是持续加血。rend也没有cool down。所以不停cle... 阅读全帖 |
|
r***u 发帖数: 1272 | 10 总的来说,DoTA是一个文化包容性很强的游戏,而且DOTA中文版翻译的还是非常非常好
的~~~
(既然这么多人看。。lz顺征人交流魔兽对战,是1v1对战喔)
后文会出现的一些常用缩写:
War3:warcraft 3 魔兽争霸3
RA:red alert 红色警戒系列
SC: star craft 星际争霸
CoD:call of duty 使命召唤
还有一些游戏的名称在文中会有注释。
好了,正文:
关于Dota和一些游戏中的GRE词汇
A字头
Abaddon :DOTA中地狱领主的名字。本意为地狱,恶魔。是圣经里的亚玻伦 (地狱的
使者)。
abomination:war3中的憎恶。abominate痛恨,憎恶。
aegis:immortal aegis 不朽盾,同时aegis是汉化版的作者。在RA2中也有神盾巡洋
舰 AEGIS Cruiser。
acolyte:侍僧,教士的助手。war3中Undead的农民。
alacrity:blade of alacrity 欢欣之刃。alacrity还有敏捷之意,所以它加敏捷属
性就... 阅读全帖 |
|
v***a 发帖数: 1242 | 11 我用多抗检测tissue中时
一般情况下都是full length的band非常清晰
cleaved form的那条短band很浅
但是单抗就只有很粗的cleaved form那一条
还有我没有很看明白你说的
多抗如果target的是胞内的那一段
那么它是怎么可以检测到cleaved form的? |
|
m*****u 发帖数: 15526 | 12 cleaved caspase 9组织核染色阳性有意义么?说明什么?因为正常对照组没有,想问
问。cleaved caspase 3 and cleaved caspase 8染色都是阴性。是上皮细胞。多谢 |
|
n**********g 发帖数: 196 | 13 OK, you want basic chemistry and here is basic chemistry.
“Since ssDNA
can not bind to the editing enzyme alone in vitro, the conclusion will be
the enzyme needs the second substance that is a conformation-ready genomic
DNA to form a functional enzyme-ssDNA-genomic target DNA complex to complete
the reaction. ”
----This is so wrong. There could be 100 reasons why an enzyme does not bind
a substance. For example, temperature, pH. Most importantly, it directly
contradicts Han’s data (Fig 1,2).
“As... 阅读全帖 |
|
c**i 发帖数: 6973 | 14 (1) Greg Sheridan, Trade and Defence: Our China line. The Australian, June
25, 2011 (availble now).
http://www.theaustralian.com.au/news/world
/trade-and-defence-our-china-line/story-e6frg6ux-1226081595229.
Quote:
"China is the strategic shadow that looms over everything today. There is a
China dimension behind US President Barack Obama's decision to pull out of
Afghanistan more quickly than his generals want. Ever since he was elected,
Obama has wanted to pivot away from the endless miseries of... 阅读全帖 |
|
c**i 发帖数: 6973 | 15 (1) Construction of China-Myanmar railway could start in December. DPA, Aug
29, 2011.
http://www.monstersandcritics.com/news/business/news
/article_1659676.php/Construction-of-China-Myanmar-railway-could-start-in-
December
Quote:
"'The whole project will take five years and cost about 20 billion US
dollars. China will bear the cost and the agreement will be based on BOT (
build, operate and transfer) for 50 years,' [Myanmar Railway Transportation
Minister] Aung Min told the German Press Agency d... 阅读全帖 |
|
w********9 发帖数: 8613 | 16 这是Shorter Oxford American English Dictionary第五版列出的与德语或者日耳曼语
相关的英语词根或词汇。
-at, suffix2. + -dom, suffix. + -ed, suffix1. + -ed, suffix2. + -en, suffix1
+ -en, suffix2 + -en, suffix5 + -en, suffix6 + -er, suffix1. + -er, suffix3
. + -er, suffix5. + -est, suffix1. + -est, suffix2. + -et, suffix2. + -eth,
suffix1. + -hood, suffix. + -ing, suffix1. + -ing, suffix3. + -ish, suffix1.
+ -kin, suffix. + -le, suffix1 + -le, suffix3 + -less, suffix. + -ling,
suffix1. + -ling, suffix2 + -ly, suffix1. + -ly, s... 阅读全帖 |
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发帖数: 1 | 17 病毒自己进化的?人工剪辑的?可以得出啥结论?
Furin is a protein that in humans is encoded by the FURIN gene. Some
proteins are inactive when they are first synthesized, and must have
sections removed in order to become active. Furin cleaves these sections and
activates the proteins.[5][6][7][8] It was named furin because it was in
the upstream region of an oncogene known as FES. The gene was known as FUR (
FES Upstream Region) and therefore the protein was named furin. Furin is
also known as PACE (Paired basic Amino ... 阅读全帖 |
|
or 发帖数: 720 | 18
I love it.
Somehow it reminded me of pride and prejudice,
not the name of mark darcy and mr. darcy but storyline and romance.
Herione was repulsive to darcys at the beginning:
bridget by darcy's diamond-shape sweater and nerdy conversation,
while liz by darcy's arrogance.
Instead they had a crush on unworthy attrative guy--
cleave and wickham respectively, who in the past impaired darcy's family:
cleave had slept with mark darcy's wife while wickham had seduced
mr. darcy's sister.
Then family d |
|
J*******g 发帖数: 8775 | 19 九
天主、我的天主,这时我经受了多少忧患、多少欺骗!当时对童年的我提示出正当生活
是在乎听从教诲,为了日后能出人头地,为了擅长于为人间荣华富贵服务的词令。因此
,我被送进学校去读书,那时我还不识读书的用处,但如果读得懈怠,便受责打。大人
们都赞成这种办法,并且以前已有许多人过着这样的生活,为我们准备了艰涩的道路,
强迫我们去走,增加了亚当子孙的辛劳与痛苦。
但是,主,我们也碰到了向你祷告的人,从他们那里,我们也尽可能地学习到、从而意
识到你是一个伟大人物,你虽则未尝呈现在我们面前,却能倾听我们、帮助我们。因为
我在童年时已开始祈求你,作为我的救援和避难所,我是滔滔不绝地向你呼吁,我年龄
虽小却怀着很大的热情,求你保佑我在学校中不受夏楚。每逢你为了我的好没有听从我
时,大人们、甚至决不愿我吃苦的父母们都笑受扑责:这在当时是我重大的患难。
主啊,是否有人怀着如此伟大的精神,以无比的热情依恋着你,我说,是否有人——因
为有时由于愚昧无知也能到此地步——虔诚依恋着你,抱着宏伟的毅力,身受世界上谁
都惊怖战栗、趋避惟恐不及的木马刑、铁爪刑等楚毒的刑罚,而竟处之泰然,甚至还热
爱着战慑失色的人们,... 阅读全帖 |
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n***w 发帖数: 2405 | 20 我不是这个领域的,读过一些AD的文章。我是这么想的:
1.因为用的是人的mutation,抗体用rodent raise,无所谓,用人的mutation就是为了
保证这个研究和临床的correlation。如果用rodent做,虽然表型一样,但是gene在
rodent上能成,但是在人上却不行,成不了大事。
2.我没有觉得主流觉得机制偏向Ca overload,这只是人们用来解释Abeta cascade的一
个思路。你找几个新的review来看看。
3.我觉得A beta产生就是要被cleaved,只是在哪里cleaved的问题。sAPP有paper说是
neurotrophic和neuroprotective作用的。 |
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s******y 发帖数: 28562 | 21 用来测proteaasome activity 的 ub-GFP必须是那种不能被cleave 的ub,
否则表达过程中就被抢先cleaved 了。
所以这个de-ub 不是一个问题。
成问题的主要原因是因为ubiquitinylated protein 也能成为lysosome,
autophagosome的底物,所以不是那么的特异。
如果这个问题对你们非常重要的话,我建议你用三种方法来重复:
1. uncleavable ub-GFP
2. P53 or H1f, by using cycloheximide or pulse trace
3. partially purified proteasome with artificial substrate
然后再加上一个加beta-lactone 的逆对照
autophagy |
|
s*******2 发帖数: 598 | 22 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 |
|
s*********s 发帖数: 110 | 23 sina上有个comment
NIBS-王晓东实验室发现Necrosis的分子机制 此博文包含图片 (2014-04-12 14:
41:15)[编辑][删除]转载▼
NIBS-王晓东实验室发现Necrosis的分子机制
NIBS-王晓东实验室发现Necrosis的分子机制
在April 10发表的Molecular Cell杂志封面文章中,王晓东实验室描述了MLKL的磷酸化
是如何导致了细胞程序化坏死的分子机制。MLKL的磷酸化使其从单体状态向多聚体状态
的转化。多聚化的MLKL可以结合多种磷脂,从而由细胞质(弥散分布)转移到细胞膜和
细胞器膜上;并在这些膜结构中形成通透性孔道,从而破坏膜的完整性,引起细胞坏死
。这一点有点类似于细胞凋亡中的重要的蛋白Bax和Bak, 在细胞凋亡领域,Bax/Bak在
细胞凋亡过程在线粒体表面聚集,多聚之后形成孔道,从而释放细胞色素C(Cyto C),
诱导细胞凋亡小体,从而激活Caspase3,最后导致细胞凋亡。
这篇文章同时介绍了王晓东实验室还开发了(Epitomics公司)能特异识别人源p-MLKL磷
酸化的单克隆抗体,并证明这一磷酸... 阅读全帖 |
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s*****g 发帖数: 7857 | 24 cleaved caspase 9阳性应该是凋亡刚开始吧,也许还没有到cleaved caspase 3的时间点
细胞方便就做个flow cytometry看看。
另外, PARP也可以看看。 |
|
s****l 发帖数: 10462 | 25 有人愿意讨论这里面提及的测序技术吗?优缺点,可行性,第一手经验等等
这里面谁现在是赢家很清楚,但是可见的将来和更远的将来谁会赢?
http://cen.acs.org/articles/92/i33/Next-Gen-Sequencing-Numbers-
全文拷贝如下
试试看图能不能贴上
/1407973177171.jpg
/1408028636124.jpg
OVER STORY
Next-Gen Sequencing Is A Numbers Game
As technical and cost barriers fall, instrument firms move their systems
into research and clinical markets
By Ann M. Thayer
Department: Business
Keywords: instrumentation, gene sequencing, diagnostics, genomics, cancer
[+]Enlarge
09233-cover-Openercxd_17647969-690
A... 阅读全帖 |
|
发帖数: 1 | 26 An alternative novel tool for DNA editing without target sequence limitation
Shu Xu†, Shasha Cao†, Bingjie Zou†, Yunyun Yue†,
Chun Gu, Xin Chen, Pei Wang, Xiaohua Dong, Zheng Xiang, Kai Li, Minsheng
ZhuEmail author, Qingshun ZhaoEmail author and Guohua Zhou
†Contributed equally
Abstract
Engineered endonucleases are a powerful tool for editing DNA. However,
sequence preferences may limit their application. We engineer a structure-
guided endonuclease (SGN) composed o... 阅读全帖 |
|
H*****9 发帖数: 11 | 27 I now discuss some of the Chinese Classical literature and prep the patient
mentally:
The Huang Di Nei Jing states that the organs produce Qi and that the Qi of
the
different organs have differing functions. However, in healthy people the Qi
of
each organ feeds and balances the receiving organ. This Qi must then travel
from
the Organs to nourish the ends of the fingers, nose, toes, and brain. So, we
have this simplified idea of Qi from the Huang Di Nei Jing. But the magic
questions are, “What is... 阅读全帖 |
|
d****y 发帖数: 2180 | 28 ☆─────────────────────────────────────☆
zhu84518 (lot) 于 (Wed Aug 8 22:41:34 2012, 美东) 提到:
A previously healthy 32-year-old woman who works as a nurse comes to the
emergency department because of a 3-week history of episodes of dizziness,
nausea, and profuse sweating that resolve with eating. She does not smoke.
She drinks four glasses of wine weekly. Vital signs are within normal limits
. Physical examination shows no abnormalities. While in the emergency
department, she becomes dizzy and... 阅读全帖 |
|
P*****s 发帖数: 375 | 29 well, I'm a newer at bichemistry, so just discuss this rather than give
an answer.
I think they just define endonuleases as those -ases that can cleave
DNA at target sites. This is a must for the insertion of intron. So
there seems no way to clone gene without it. (Or you may ask: Can a
gene be cloned without cleaving DNA?) |
|
s****n 发帖数: 174 | 30 还是老外呢。怎么,难道还怕牧师忘了,给他递纸条 :)
版本的确很多,大同小异,这是其中一个:
Do you WSN, take LD, whom you hold by the hand, to be your true and lawful
wife, to love and cherish her, in joy or pain, in sickness or health and
forsaking all others, to cleave to her only, so long as you both live? (NOTE
Do you LD, take WSN, whom you hold by the hand, to be your true and lawful
husband, to love and cherish him, in joy or pain, in sickness or health and
forsaking all others, to cleave to him only, so long as you both live? ( LD
h |
|
s*****5 发帖数: 31 | 31 English channel carved by huge ancient flood
Ian Sample, science correspondent
Wednesday 18 July 2007 09.26 EDT
A cataclysmic flood cleaved Britain from France hundreds of thousand years
ago, in a violent act of nature that carved out the white cliffs of Dover
and set the course of history for a new island.
High-resolution sonar images of the English channel collected over more than
20 years have revealed a deep scar in the seabed that was ripped from the
limestone bedrock by a torrent of water ... 阅读全帖 |
|
h******i 发帖数: 21077 | 32 No no no.
Blunt weapon (such as stone axe) can't "chop", it could only "blow" or "
knock".
Pangu was "Chopping" (or "cleave") the world into sky and earth, so he or
she was using a chopping axe, not blunt axe or stone axe.
Unless you could prove that Pangu was "knocking" the world into two parts. |
|
c*****a 发帖数: 1238 | 33 Bt protein: Cry1Ab, one of Cry proteins.
The midgut of these insects is alkaline, which allows for the activation of
the Cry proteins once the proteins are ingested.
The Cry proteins are then “dissolved into protoxins, which are cleaved by
proteases into active toxins".
The activated proteins then bind to cells in the insect’s midgut and form
pores in the cells’ outer membranes.
This leads to the swelling and destruction of midgut cells . The digestive
systems of the insects are then paralyzed |
|
R****a 发帖数: 6858 | 34 美国为何就华为进军加拿大发出警告
收听 (04:52) 录入个人播放列表 下载 Embed
作者 蒙特利尔特约记者 潘卫
5月15日,前美国反间谍机构负责人克利芙(Michelle K. Van Cleave)向加拿大发出
警告,指它允许中国华为公司参与境内大型电信工程的做法,将把加拿大和美国的高速
网络置于中国网络间谍的控制之中,进而威胁整个北美地区的安全。克利芙在美国首都
华盛顿接受了加拿大广播公司的专访,她强调美国和澳大利亚早已把华为视为一个潜在
的安全威胁,把它排斥在大型电信项目之外,例如澳大利亚政府曾禁止华为投标其“国
家宽带网”,加拿大情报机构也曾就此发出过警告。
克利芙显然是对华为近来在加拿大重要电信项目上取得突破表示不满,今年2月,华为
宣布将承建加拿大3G骨干网络,它和加拿大两家最大的电信运营商Telus和贝尔移动签
署了合同,为它们提供LTE网络升级,并提供全加拿大各地的LTE无线接入网设备。在2
月份与贝尔公司签署项目合同时,当时正在访华的加拿大总理哈珀还出席了在北京的签
字仪式。
克利芙的警告在加拿大各界引起震动,两天后,总理哈珀向提出质询的国会议员们保证
,加... 阅读全帖 |
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w*********g 发帖数: 30882 | 35 英国夫妇给宝宝哼唱《粉红猪小妹》主题曲,被穆斯林赶下公交车(转载)
楼主:chai15221 时间:2014-09-27 21:44:00 点击:22277 回复:391
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有消息披露,一对夫妇给他们的自闭症女儿哼唱《粉红猪小妹》主题曲被视为种族
主义者,因而在途中被赶下公车。来自罗瑟勒姆(Rotherham)的尼克·巴恩菲尔德(Nick
Barnfield)和莎拉·克里夫斯(Sarah Cleaves)带着他们的女儿坐在一辆从设菲尔德开
往唐克斯特的公车上,忽然,15个月大的女儿开始哭闹。夫妇俩为了逗女儿开始哼唱一
首歌,但另一名乘客称他们种族主义者歧视她,驾驶员随后要求他们下车。夫妇俩称一
位戴着头巾的妇女攻击他们的哼唱声,她认为他们在影射伊斯兰教的猪肉禁忌。女子向
公车驾驶员投诉,司机竟然随后要求这对夫妇在离家还有两英里远的地方下车,这样事
情会“大事化小,小事化了”。24岁的巴恩菲尔德说:“我们非常尴尬,感到非常羞辱
,很生气,我们没做错什么事,就是想逗我们的小女儿,但这些人觉得我们过错很大,
太羞辱人了。一位妇女走上... 阅读全帖 |
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发帖数: 1 | 38 Abstract
We show here that, unlike most other prokaryotic Argonaute (Ago) proteins,
which are
DNA-guided endonucleases, the Natronobacterium gregoryi-derived Ago (NgAgo)
can
function as a DNA-guided endoribonuclease, cleaving RNA, rather than DNA,
in a
targeted manner. The NgAgo protein, in complex with 5’-hydroxylated or 5’-
phosphrylated oligodeoxyribonucleotides (ODNs) of variable lengths, split
RNA targets
into two or more fragments in vitro, suggesting its physiological role in
bacteria and... 阅读全帖 |
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