x*****7 发帖数: 7326 | 1 【 以下文字转载自 Military 讨论区 】
发信人: xiaxie7 (虾蟹大将), 信区: Military
标 题: 河北部队的病毒感染事件
发信站: BBS 未名空间站 (Wed Feb 29 01:30:45 2012, 美东)
本虾认为可能是某种重组病毒感染。
野生型的腺病毒(adenovirus)不会这么厉害。
考虑到那个部队的官兵是去广东训练染上的,本虾认为目前不能百分之百排除SARS。 |
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T******J 发帖数: 829 | 2 刚刚买了一个Basset Hound PUPPY,9个星期。(上照) Breeder说在8个星期时已经打
了DE-WORM,PAPERWORK上海有两个STICKER, 一个是CANINE CORONAVIRUSVACCINE, 另
一个是CANINE DISTEMPER-ADENOVIRUS TYPE2-PARAINFLUENZA-PARVOVIRUSVACCINE。
不知道是否都需要打?
下个星期打算去CHECK VET。
谢谢!! |
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M****e 发帖数: 70 | 3 in the case of a couple of start codons, the context arround
is important to determine the initiation site. and the most
famous one is found by Kozak. basically, leaky scanning means
that ribosome scans the mRNA and bypasses the first start codon
it meets and initiates from a downstream one. some virus proteins
utilizes this mechanism for translation initiation.
shunting means "jumping". in adenovirus and hepatitis virus,
some genes use such ribosome shunting mechanism for translation
initiation |
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m*****i 发帖数: 29 | 4 Check out this paper:
Xu ZL, Mizuguchi H, Mayumi T, Hayakawa T.
Regulated gene expression from adenovirus vectors: a systematic comparison of
various inducible systems.
Gene. 2003 May 8;309(2):145-51.
Actally I wasted half year on Clonetech's tet-on/off system. It is very leaky
so that in some cell types (MEF is worst) you can never control the basal mRNA
expression level at all. After half year's work, I called the campany and also
found out this paper that compared different commercial inducib |
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C****1 发帖数: 89 | 5 新手我最近苦于纠结一个关于IP实验的问题,特发此帖向大家求教。
我的目的蛋白A如果在肿瘤细胞(此蛋白已证实该细胞里正常水平不表达)里过度表达
会含有FLAG tag(通过adenovirus vector infection)。我用此蛋白来pull down 细胞
里的另外一个蛋白B。我用FLAG 抗体IP, Western blot 检测此两种蛋白A和B。结果在
正常对照的样品发现anti-Flag 抗体也能非特异性的结合蛋白B,即在blot也能检测到
B的条带。
请问如何消除抗体非特异性结合其他蛋白,还是我在那些方面需要改进?谢谢。 |
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T**********r 发帖数: 287 | 6 如果是lentivirus或adenovirus呢 |
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h******o 发帖数: 23 | 7 我有少量的包装好的病毒颗粒,需要用293细胞复制我的病毒颗粒。
我发现第一步病毒颗粒接触细胞时,是要用serum free medium。
不知道板上的高人可否指点一下,serum 是不是有抑制作用呀?可否直接用普通的
medium?
多谢! |
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r****r 发帖数: 379 | 8 这个问题比较tricky,一般来说,因为血清里的一些成分,比如补体,免疫球蛋白等会结合病毒,因此干扰细胞对病毒的吸收。但这依病毒类型而定,有些就不受影响。
但也因为如此,有些不含CAR的细胞,相反必须依靠补体之类的东西才能吸收这些病毒,所以对这些细胞的侵染还必须加血清。 |
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R*****w 发帖数: 447 | 11 Thank you very much, sunnyday!
I figured it out and found that pcDNA3.1 does have sv40 origin.
The following is some information from Wiki, in case someone else is
interested in this:
HEK 293 cells were generated in early 70s by transformation of cultures of
normal human embryonic kidney cells with sheared adenovirus 5 DNA in Alex
Van der Eb's laboratory in Leiden, Holland.
An important variant of HEK293 cell line is the 293T cell line that contains
, in addition, the SV40 Large T-antigen, that |
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T**********r 发帖数: 287 | 12 现在正在用invitrogen的viralpower adenoviral expression system, infect 293a
cell, 在37度培养箱培养。
箱内一层共有16个15cm plate,其中十个是infected的,其它6个是未被侵染的,用来
split
cell的。3天后发现所有的plate都被侵染了,都表达了mCherry.
可以肯定的一点绝不可能是我不小心将virus加到其它6个plate里了。会不会virus自
己挥发出来
侵染了其它盘,如果这样,应该怎样防护。希望大家分析一下,感激不尽! |
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D******9 发帖数: 2665 | 14 I think it could be contaminated in the cell culture wood. |
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r*******w 发帖数: 127 | 15 The same happened to my experiments. The uninfected plates maybe got
contanminated somwhere (in hood or in incubator). When uninfected plates and
infected plates were seperated to culture in different incubators, the
uninfected plates kept normal.
293a |
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w***e 发帖数: 269 | 16 adenovirus 侵染做liver gene knockdown 比较容易.其他tissue 难点.你的基因是在
liver里面么? |
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T**********r 发帖数: 287 | 17
我是往pancreas里注射,让adenovirus侵染pancreas,并且在pancreas里表达,这个技
术问题已
经解决了。我是想知道pAd/CMV/mCherry-linker-shRNAmir这样的设计是否合理。
linker是
polycistronic的2A。多谢答复。 |
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p*****y 发帖数: 44 | 18 我是往pancreas里注射,让adenovirus侵染pancreas,并且在pancreas里表达,这个技
术问题已经解决了
能讲讲这个技术吗?把老鼠打开来直接注射? |
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r***e 发帖数: 2539 | 19 Thank everyone for the discussion!
As pineseed pointed out, too many topics were discussed here.
But my personal interests are:
1. what is the cell origin of different type of lung cancers?
2. does cell origin or mutation decide tumor types?
3. how to model different type of lung cancer?
HGTPase, it seems you are working in this field (more lung development than
lung cancer?), and I hope you can give your opinions. We can also discuss in
private mails for technique details.
1. ... 阅读全帖 |
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b*********8 发帖数: 116 | 20 如果只能用transfection的话,哪位做过用lipofecamine的?或者只能用electro
多谢!! |
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w********r 发帖数: 1431 | 22 你的primary myocyte是说satellite cell么?
如果是的话,我用lipoRNAiMax往primary myoblast里转过siRNA,效率还可以,我做
western看过。
转plasmid确实是进不去,我都是用Adenovirus。
不过看业内大牛的paper也有往里面裸转质粒的,具体用的啥试剂记不清楚了,你可以
去查查Michael Rudnicki的paper. |
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j******i 发帖数: 939 | 23 Gamma-retrovirus and lenti are both efficiently transduced into cells, but
gamma-retro can not infect non-dividing cells. Both Gamma-retro and lenti
insert into genome and thus have insertional mutagenesis risk, however,
gamma-retro usually inserts into transcription start sites and more likely
to disrupt genome gene expression. Lenti virus, however, is sort of safer
virus with no preference to start point while inserting.
Adenovirus is episomal virus and can only infect dividing cells. This mea... 阅读全帖 |
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m****r 发帖数: 383 | 24 把Ad-Cre打到mammary gland of a Cre reporter mouse,理论上会诱导gfp的表达,只
是不知道从注射,到cre-mediated recombination发生,直至gfp蛋白表达,这个
过程需要多久?一周? |
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w*e 发帖数: 740 | 25 will not take that long. |
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D*a 发帖数: 6830 | 26 请问你有管用的protocol么?我正好想弄一个recombinaison的老鼠出来的说 |
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D*a 发帖数: 6830 | 27 PS 打算注射到testis里面,拿后代的KO。不知道可行否? |
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w*e 发帖数: 740 | 28 I performed CAG-CREER+; R26-EYFPloxp+
by injecton TMX in vivo, we can see EYFP within 72hs
therefore, I say by adcre will not cause 1 week |
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h********n 发帖数: 4079 | 30 Who has done intra-ductal injection of virus into mouse mammary gland?
Thank you. |
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m****r 发帖数: 383 | 31 Thanks for the info. May I know what tissue(s) you checked? |
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m****r 发帖数: 383 | 32 looks like an unrelated paper... but thank you! |
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s*******b 发帖数: 1012 | 34 adenoviral construct已经有了,想找个公司做成virus. 以前没做过viurs,毫无经验
,还希望
版上的前辈指点一下,推荐几个有亲身经验的公司,好跟老板交差,多谢多谢。
By the way,实验室在西海岸 |
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A******y 发帖数: 2041 | 36 There are a lot of immortalize techniques...for example the most common
human kidney cell line Hek293 is an embryonic kidney cell transformed with
adenovirus 5 DNA. Usually the immortalization is included in the cell line
information sheet.
http://www.atcc.org/ATCCAdvancedCatalogSearch/ProductDetails/ta |
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g*********d 发帖数: 233 | 37 MicroRNA 和病毒
作者:王磊
上海交通大学生命学院研究生
摘要:
microRNA(miRNA)是真核生物体内一组内源性的 21—25nt 长的非编码蛋白质的短序列
RNA,它们能通过碱基配对的方式与生物体中的 mRNA 分子相结合来参与调节基因的表
达。自 2000 年至今,miRNA 的研究发展的速度极快并且影响的范围也非常的广泛,而
且分别在2002年和2003年度中miRNA入选了Science杂志年度十大科技突破。因此,
miRNA是 RNA 研究的又一个突破,被称为 RNA 的第二次革命。目前的研究已发现
miRNA 参与了很多重要的生命活动过程涵盖了发育、凋亡、代谢、人类疾病以及病毒侵
染等方面。在病毒侵染宿主和宿主抵抗病毒的过程中病毒编码的 miRNAs 和宿主编码的
miRNAs 均发挥了重要的作用。这无疑为病毒的侵染以及宿主抵抗病毒的侵染的研究提
供了新的思路和新的领域。因此,本文的主要目的在于阐述 miRNA 的一些基本概况以
及在病毒中主要的研究方向和进展,最后对这个具有跨时代意义的小分子进行展望。
关键词:miRNA vmiRNA 功能 宿主的 miR... 阅读全帖 |
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D***a 发帖数: 516 | 39 两者之间有关系吗?为什么都有adeno这个词缀?好多癌症为什么都叫adenocarcinoma? |
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D***a 发帖数: 516 | 40 哦,那就是一个是从腺体中分离的,一个是分泌腺组织癌变,没什么本质关联。
wikipedia |
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b******n 发帖数: 4225 | 41 1. Automated mass immunization of poultry: the prospect for nonreplicating
human adenovirus-vectored in ovo vaccines.
Avakian AP, Poston RM, Kong FK, Van Kampen KR, Tang DC.
Expert Rev Vaccines. 2007 Jun;6(3):457-65. Review.
2. DNA vaccines for poultry: the jump from theory to practice.
Haygreen L, Davison F, Kaiser P.
Expert Rev Vaccines. 2005 Feb;4(1):51-62. Review.
Email: s****[email protected]
Thanks! |
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q**********0 发帖数: 335 | 42 For example, HEK293 was transformed by Adenovirus protein. |
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g****y 发帖数: 101 | 43 To Carwash: Thank you very much for comment
I can't agree more with you about the low efficiency in infecting the T
cells in vivo with virus. That is why I mentioned the potential problem 4(
AAV载体的靶向问题).
As you suggested, we could modify the virus caspid, for example, mutate the
liver cell recognition sequence and express a chimeric protein combining the
AAV capsid protein and the T cell ligand so that we can increase the
infection efficiency in T cells.
Do you think this approach works?
Here ar... 阅读全帖 |
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F******p 发帖数: 2099 | 44 The commercial lab virus does not propogate unless co-infect with helper
virus.
adv does infect human, the symptom is like getting a cold. Normally your
immune system will kill the virus without problem, but no idea about
pregnancy.
retrovirus infect human too. |
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w******n 发帖数: 352 | 45 看你们有没有自己插入什么基因,本身病毒是没有问题,但大多数实验室用来携带基因
,这些基因会有问题。
但学校不会管,因为危害是理论上,实际不好界定。你自己小心就是了,不要闹到老板
那里,找不必要的麻烦。 |
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s*****g 发帖数: 7857 | 46 非用什么鬼病毒?我当初连做病毒细菌的实验都不沾.
我就电转.别人不好转而用病毒转的,我都用电转.一样成功.而他们的病毒转的后来转的
基因后来都消失了,而我电转,抗生素筛选的还好好的. |
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h****s 发帖数: 40 | 47 天才呀!
佩服得很呢!
全世界能做到您这一点的我觉得没有第二个人了。 |
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f******l 发帖数: 101 | 48 和我在网上查到的信息差不多。实验用应该不会自己复制,但是可以infect人。
我们现在用了好几种基因,包括p53,E6,Rb,E7,TRCP和Sleeping beauty transposase 。所以还是有点担心。
谢谢stilconfused,Fatsheep和waterman。 |
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f******l 发帖数: 101 | 49 我以前自己的project也是用电转做stable cell line的。
但是现在最主要是工作以后protocol不由自己选择了,二是现在是transient,电转死
的细胞太多,而普通的transfection reagent效率又太低。 |
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j****x 发帖数: 1704 | 50 载体本身是很安全的,如果是在合乎生物安全防护标准的实验室条件下按照标准操作程
序进行实验,风险很小,没有什么可担心的。假如出现意外例如你梦中的情形,可以要
求同事帮助清理你尽快离开现场并向实验室生物安全主管汇报,被感染的几率也是非常
的小,何况你体力很可能早已存在针对腺病毒的抗体了 |
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