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2018 - 06 - 08
2018年6月6日讯 /生物谷BIOON /——PERIOD 2 (PER2)蛋白的多点磷酸化是决定哺乳动物生物钟周期的关键步骤。过去的研究认为PER2被酪蛋白激酶1(CK1)磷酸化的过程需要一种未被发现的蛋白激酶来启动,而CK1是一种从藻类到人类都高度保守的生物钟必需蛋白。这些磷酸化过程可以稳定PER2,延缓其降解以延长生物钟周期。图片来源:CCO public domain为了找到这个必需的蛋白质,一个由杜克-新加坡国立大学医学院研究人员领导的国际团队对小鼠PER2启动磷酸化的过程进行了综合的生物化学和生物物理学分析,结果惊讶地发现CK1δ/ε就是他们寻找的启动激酶。此外,他们还发现CK1ε和一个最近发现的CK1δ2剪接变体可以比CK1δ1更有效地启动磷酸化。尽管过去研究显示CK1对PER2的磷酸化对细胞环境很不敏感,但是研究人员通过使用生物节律磷酸化开关数学模型发现CK1羧基端尾部使得生物钟周期对细胞信号敏感。这些研究表明CK1羧基端是生物节律的一个重要的调节因子。(生物谷Bioon.com)参考资料:Rajesh Narasimamurthy et al, CK1δ/ε protein kinase primes the PER2 circadian phosphoswitch, Proceedings of the National Academy of Sci...
2
2019 - 03 - 22
给你一个冰淇淋和一块蛋糕,你先吃哪个?一块面包和一个牛油果呢?两个一块吃吗?1941 年微生物学家 Jacques Monod 给细菌提出了同样的问题。他给大肠杆菌同时提供两种食物(碳源),结果看到了两类不同的现象:对某一些食物组合(比麦芽糖和苹果酸),细菌两种同时吃,而对另一些组合(比如葡萄糖和乳糖),细菌要先吃完一种(葡萄糖),然后再会去吃另一种(乳糖)。随后 Monod 和 François Jacob 一道,找出了细菌在有葡萄糖和乳糖的情况下只吃葡萄糖的分子机制—乳糖操纵子:在这两种糖都存在的时候,细胞不会去表达运输和消化乳糖的相关基因,并获得了 1965 年的诺贝尔生理学奖。可是为什么细菌要挑食呢?为什么对于有些食物的组合它们又不挑呢?北京大学汤超教授团队发现细菌的这一套进食策略能够使它们最优化的生长。如果哪个菌种想任性一把,同时吃葡萄糖和乳糖,或者对麦芽糖和苹果酸挑食,那么它们的代价就是生长速率的降低,进而面临被淘汰的危险。结合代谢网络的拓扑特征和细胞内蛋白资源的优化分配,研究团队的理论模型定量的解释了细菌面对多种碳源的进食策略。在两类碳源同时被细菌消耗的情况下,模型预测并用实验验证了细菌内不同氨基酸上的碳原子来自两种碳源的比例。这个例子进一步说明了在看似复杂的生命现象中,存在简洁的原理与定量的规律。相关文章发表在《自然 - 通讯》杂志上。北京大学定量生物学中...
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2019 - 07 - 26
科技日报北京 7 月 30 日电 (记者付丽丽)“建立严格的法律责任制度,对违反《疫苗管理法》规定构成犯罪的,依法从重追究刑事责任,货款不足 50 万元的按 50 万元计算。”30 日,在国家药监局主办的 “《疫苗管理法》颁布法规解读宣贯研讨会上”,该局政策法规司副司长吴丽雅说。《疫苗管理法》是全球首部综合性疫苗管理法律。具体来讲,吴丽雅介绍,生产、销售的疫苗属于假药的,没收违法所得和违法生产、销售的疫苗以及专门用于违法生产疫苗的原料、辅料、包装材料、设备等物品,责令停产停业整顿,吊销药品注册证书,直至吊销药品生产许可证等,并处违法生产、销售疫苗货值金额 15 倍以上 50 倍以下的罚款。生产、销售的疫苗属于劣药的,处违法生产、销售疫苗货款金额 10 倍以上 30 倍以下罚款。吴丽雅为读者算了一笔账。生产、销售的疫苗属于假药的,最低罚款:50 万元 ×(15 倍至 50 倍)=750 万元至 2500 万元;生产、销售的疫苗属于劣药的,最低罚款: 50 万元 ×(10 倍至 30 倍)=500 万元至 1500 万元。疫苗是特殊的药品,事关公共安全。吴丽雅表示,长春长生问题疫苗事件暴露出我国在疫苗监管方面存在主体责任不落实、质量安全管理不到位、职业化专业化监管力量薄弱等问题。鉴于此,《疫苗管理法》明确提出国家对疫苗实行最严格的管理制度,对疫苗的研制、生产、流通、...
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产品名称:

Premo™ FUCCI Cell Cycle Sensor (BacMam 2.0)

上市日期: 2019-04-18
产品类别: INVITROGEN

规格

Color:Green, Red-Orange
Delivery Method:Baculovirus (BacMam System)
Detection Method:Fluorescent
Emission Class:Visible
Excitation Class:Visible
Excitation⁄Emission (nm):488⁄510 (Green), 555⁄584 (Red)
For Use With (Equipment):Flow Cytometer, Fluorescence Microscope, Fluorescent Imager, High Content Instrument, Tali® Image-Based Cytometer
Form:Liquid
Label or Dye:GFP (EmGFP), RFP (TagRFP)
Product Line:Premo™
Product Size:2 x 200 µL
Quantity:2 vials (200 µl each)
Sub-Cellular Localization:Nucleus
Technique:Fluorescence Intensity
Shipping Condition:Wet Ice


  • 产品描述
  • 产品功能
  • 产品参数

描述

Premo™ FUCCI Cell Cycle Sensor is a fluorescent, two-color sensor of cell cycle progression and division in live cells. It allows accurate and sensitive cell cycle analysis of individual cells or a population of cells by fluorescence microscopy, flow cytometry, or high-content imaging.

At 2 x 200 µL, this is a smaller pack size version (1/5 volume) of the popular full-size product and perfect for testing its performance in your system.

The Premo™ FUCCI Cell Cycle Sensor is delivered by highly efficient BacMam 2.0 technology, enabling cell cycle studies in essentially any cell type. Provided in a ready-to-use format—simply add, incubate, and image. Premo™ FUCCI affords highly efficient transient expression in cell lines, primary cells, and stem cells.

Premo™ FUCCI Cell Cycle Sensor is:

Accurate: Cell-cycle controlled expression of bright GFP and RFP indicators for live cell analysis of individual cells or populations
Highly Efficient: >90% transduction of a wide range of mammalian cell lines, including primary cells, stem cells, and neurons
Fast and Convenient: Simply add Premo™ FUCCI Cell Cycle Sensor to your cells in complete medium, incubate overnight, and analyze
Safe: BacMam reagents are non-integrating and non-replicating in mammalian cells, lack observable cytopathic effects, and are suitable for biosafety level (BSL) 1 handling

Study Cell Cycle in the Relevant Cellular Context
If you are studying cell cycle regulators, cell differentiation, or are developing anti-cancer compounds against cell cycle-related targets, Premo™ FUCCI is a superbly sensitive and accurate sensor tool. The efficient transduction and transient expression allows you to conduct these experiments in the biologically relevant cell types.

Simple, Quick Cell Cycle Visualization
Simply add the ready-to-use Premo™ FUCCI cell cycle sensor to your cells in complete medium, incubate overnight, then visualize cell cycle progression in populations of cells using fluorescence microscopy, flow cytometry, or high content imaging instruments. Cells change from red in the G1 to yellow in the G1/S interphase and green in S, G2, and M phases, as fusions of emGFP and TagRFP coupled to two cell cycle-regulated proteins are expressed and degraded.

How It Works
Premo™ FUCCI is derived from the fluorescence ubiquitination cell cycle indicator (FUCCI), developed by Miyawaki and colleagues (Ref 1). FUCCI is based on two cell cycle-regulated proteins, geminin and Cdt1, fused to one green (emGFP) and red (TagRFP) fluorescent protein, respectively. As Cdt1 and geminin are present only during specific phases of the cell cycle, the fluorescent protein partner is similarly cell-cycle dependent. Ubiquitination by specific ubiquitin E3 ligases target the fusion constructs in the proteasome for degradation. E3 ligases display temporal regulation of activity, resulting in biphasic cycling of geminin and Cdt1 levels during the cell cycle. Geminin-GFP is degraded in the G1 phase; the presence of Cdt1-TagRFP is indicated by red fluorescence within nuclei. During the S, G2, and M phases, however, Cdt1 is degraded and only geminin-GFP remains, giving cells green-fluorescent nuclei. During the G1/S transition, when Cdt1 levels are decreasing and geminin levels are increasing, both proteins are present, giving a yellow-fluorescent nuclear signal. This dynamic color change (red to yellow to green) clearly displays progression through the cell cycle and division.

Powered by BacMam 2.0 Technology
BacMam technology is based on an insect virus (baculovirus) for efficient transduction and transient expression in mammalian cells. The Premo™ FUCCI cell cycle sensor uses BacMam 2.0 technology to enable cell cycle studies in essentially any cell type, from cell lines to stem cells by incorporation of a mammalian expression cassette for the FUCCI construct. Compared with BacMam 1.0 technology, BacMam 2.0 does not require the use of an enhancer and can be used in complete medium. There is no need to purify plasmid or worry about vector integrity and quality. No lipids, dye-loading chemicals, or other potentially harmful treatments are required. BacMam 2.0 incorporates elements that greatly enhance transduction efficiency and expression levels: a pseudotyped capsid protein for more efficient cell entry, an enhanced CMV promoter, and the Woodchuck hepatitis post-transcriptional regulatory element (WPRE).

Titratable Expression that Lasts Up to Two Weeks
The transient transgene expression lasts from about 5 days in transformed cell lines to up to two weeks in slowly dividing cells, such as some primary cell types; we have imaged terminally differentiated neurons more than three weeks after transduction. In addition, BacMam technology permits defined optimization because it gives you the ability to precisely titrate expression levels. Baculoviruses do not replicate in mammalian cells, nor are viral genes expressed, giving them an excellent safety profile and lack of cytopathic effects on mammalian cells.

Learn more about the Premo™ FUCCI Cell Cycle Sensor

Reference:

1. Sakaue-Sawano A, Kurokawa H, Morimura T, Hanyu A, Hama H, Osawa H, Kashiwagi S, Fukami K, Miyata T, Miyoshi H, Imamura T, Ogawa M, Masai H, Miyawaki A. Visualizing spatiotemporal dynamics of multicellular cell-cycle progression. Cell. 2008 Feb 8;132(3):487-98. PubMed PMID: 18267078

Related products
We offer a range of BacMam-based reagents beyond CellLight® reagents, including the BacMam GFP Transduction Control that is ideal to test out the technology and optimize transduction conditions, Premo™ Biosensors, including Premo™ Autophagy Sensor, ion channel drug targets, pathway analysis kits, and more.

Learn more about these products and the BacMam technology

See other imaging tools and reagents

For Research Use Only. Not intended for any animal or human therapeutic or diagnostic use


规格

Color:Green, Red-Orange
Delivery Method:Baculovirus (BacMam System)
Detection Method:Fluorescent
Emission Class:Visible
Excitation Class:Visible
Excitation⁄Emission (nm):488⁄510 (Green), 555⁄584 (Red)
For Use With (Equipment):Flow Cytometer, Fluorescence Microscope, Fluorescent Imager, High Content Instrument, Tali® Image-Based Cytometer
Form:Liquid
Label or Dye:GFP (EmGFP), RFP (TagRFP)
Product Line:Premo™
Product Size:2 x 200 µL
Quantity:2 vials (200 µl each)
Sub-Cellular Localization:Nucleus
Technique:Fluorescence Intensity
Shipping Condition:Wet Ice


规格

Color:Green, Red-Orange
Delivery Method:Baculovirus (BacMam System)
Detection Method:Fluorescent
Emission Class:Visible
Excitation Class:Visible
Excitation⁄Emission (nm):488⁄510 (Green), 555⁄584 (Red)
For Use With (Equipment):Flow Cytometer, Fluorescence Microscope, Fluorescent Imager, High Content Instrument, Tali® Image-Based Cytometer
Form:Liquid
Label or Dye:GFP (EmGFP), RFP (TagRFP)
Product Line:Premo™
Product Size:2 x 200 µL
Quantity:2 vials (200 µl each)
Sub-Cellular Localization:Nucleus
Technique:Fluorescence Intensity
Shipping Condition:Wet Ice


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