Luciferase refers to a group of bioluminescence producing enzymes that enable researchers to study gene expression and regulation.
光素酶是指，一群生生物冷光，使研究人能研究基因表及的酵素。
1. 光的火，在光素酶的酵素助下，照亮森林。
Glowing fireflies illuminate a forest with the help of luciferase enzymes.
Luciferase is a group of enzymes that oxidize a substrate known as luciferin to produce light. For bioluminescence, the firefly luciferase enzyme catalyzes the oxidation of D-luciferin in the presence of oxygen (O2), adenosine triphosphate (ATP), and magnesium ions (Mg2+).
光素酶是一群，氧化一被通光素之酶作用物(因酶或酵素的作用，引起化反的物)，以生光的酵素。就生物冷光而言，火的光素酶酵素，在氧(O2)、三磷酸腺苷(ATP)及子(Mg2+)存在的情下，催化D-光素的氧化。
This catalytic reaction converts the excited oxyluciferin molecule to its ground state, emitting visible light. Bioluminescence observed in various organisms may utilize different chemical processes. However, they all share a common feature: the production of light through enzyme-catalyzed oxidation reactions.
此催化反被激的氧光素分子，到出可光的基。於各生物中，被察到的生物冷光，可能利用不同的化化程。不，它全有共同特性：也就是，光的生是透酵素催化的氧化反。
Although bioluminescence has been observed for many centuries, its application in biomedicine is relatively recent. Scientists traced back the first reference of bioluminescence to the Greek philosopher Aristotle in 384-322 BCE.
然，生物冷光已被察，世。不，其在生物方面的用，相上是近代的。科家生物冷光的首次提及，追溯到公元前(Before the Christian Era)384-322年的希哲家里斯多德。
Almost three centuries later, Gaius Plinius Secundus, a Roman author, conducted an in-depth study on bioluminescence. He discovered many animals that had illuminating capacity such as purple jellyfish, mollusks, fireflies, and glowworms.
近三世後，作家Gaius Plinius Secundus，生物冷光行了深入研究。他了，多具有光能力的物，如紫色水母、物、火及光等。
French pharmacologist Raphael Dubois observed that a specific constituent of the click beetle was essential for bioluminescent reactions. He extracted two components from beetle abdomens, naming the substrate luciferine and the enzyme responsible for the reaction luciferase.
法理家Raphael Dubois察到，磕甲的一特定成分，於生物冷光反是不可或缺的。他甲腹部提取了成分，此酶作用物命名光素，而此酵素致上述光素酶反。
Over many years, scientists across the world conducted multiple experiments and established that luciferase enzymes and substrates are species-specific. For example, the Gaussia and Renilla luciferases use coelenterazine as their substrate to emit blue to cyan light, while Cypridina uses its own genus-specific luciferin to generate blue light.
多年，世界各地的科家，行了多次且，光素酶及酶作用物是物-特有的。譬如，Gaussia及Renilla光素酶，使用腔素作它出色到青色光的酶作用物，而Cypridina使用本身之-特有的光素，生光。
In 1985, scientists made a milestone scientific breakthrough by cloning the firefly luciferase (FLuc) in Escherichia coli, enabling an unlimited source for this enzyme. Different types of luciferases are present in aquatic and terrestrial species including insects, bacteria, and fungi. Scientists estimated that around 75% of sea animals are bioluminescent.
於1985年，科家藉由，在大菌中，性生殖了火的光素酶(FLuc)，得了科突破性里程碑，使其能作此酵素的一限源。不同型的光素酶存在於，包括昆、菌及真菌等，水生生物中。科家估，大75%的海洋物是生物冷光的。
In nature, a luciferin–luciferase system is present in the firefly (Photinus pyralis), bacteria (Vibrio harveyi), sea pansy (Renilla reniformis), jellyfish (Aequorea victori), ostracod (Cypridina noctiluca), dinoflagellates (Gonycaulax), and copepod (Gaussia princeps).
在自然界中，有光素光素酶物的物系存在於，火(Photinuspyralis：方火)、菌(Vibrio harveyi：哈氏弧菌)、海堇(Renilla reniformis)、水母(Aequorea victori：多利多管光水母)、介形(Cypridina noctiluca)、甲藻(Gonycaulax：膝藻)及足(Gaussia princeps：高斯王子)中。
Not all bioluminescent marine organism systems can be used for animal imaging because the majority of them generate blue light that does not penetrate tissue appropriately for detection. Furthermore, certain small molecule substrates of marine organisms are unstable and easily oxidized without luciferase.
非所有生物冷光的海洋生物物系，能被使用供作物造影之用。因，大多它生，法穿透，供作探知之用的光。此外，海洋生物的某些小分子酶作用物不定，且在有光素酶的情下，容易被氧化。
Table 1: Different types of luciferase and their applications
表1：不同型的光素酶及其用(原文)
Among the different types of luciferases, scientists use FLuc widely for biological experiments because of its high quantum yield, elevated signal to noise ratio, and magnificent color. Furthermore, these luciferases trigger yellow-green to red light emission, depending on reaction conditions, which has a higher cellular or tissue penetrance. Luciferin is biologically stable as it is not oxidized in the absence of the enzyme.
在不同型的光素酶中，FLuc因其高量子率、提高的信噪音比及的色彩，而被科家泛使用供作生物之用。此外，些光素酶，出取於反件，而具有高胞或穿透力之-到光。光素生物上是定的，因有光素酶，它不被氧化。
Biochemical engineers created several generations of FLuc that exhibit improved brightness and other properties. For example, Luc2 is a second-generation firefly luciferase with an optimized codon that facilitates its expression in mammalian systems.
生化工程造了，代展出改善亮度及其他性的FLuc。譬如Luc2是具有，在哺乳物物系中促其表，化密子之第二代火的光素酶。
The new firefly bioluminescence system Akaluciferase (AkaLuc)-AkaLumine produces greater signal strength than the standard Fluc-luciferin reaction.
新的火生物冷光物系，Akaluciferase (AkaLuc)-AkaLumine生，比Fluc光素之反，更的信。
AkaLuc-AkaLumine (named after the Japanese word for the color red, aka) is a non-invasive and highly sensitive bioluminescence system that emits red light in the near-infrared (NIR) range for better tissue penetration. Besides FLuc, researchers also use NanoLuc luciferase (NLuc) and Renilla luciferase (RLuc) for biological experiments.
AkaLuc-AkaLumine(是以日代表色的名)是一，在近外(NIR)出光，供作更佳穿透之用，非侵入性、高度敏感的生物冷光物系。除了FLuc之外，研究人也使用，NanoLuc光素酶(NLuc)及Renilla光素酶(RLuc)，供作生物之用。
Each luciferase type has different advantages and disadvantages depending on the application. For instance, both FLuc and RLuc have short protein half-lives making them favorable transcriptional reporters. Even though FLuc is brighter than RLuc, the former is more vulnerable to enzyme inhibition.
取於用，每一光素酶具有不同的及缺。例如，FLuc及RLuc 者皆具有短的蛋白半衰期，使它成良好的信息物。即使，FLuc比RLuc更亮。不，前者容易受到酵素抑制。
Because FLuc and RLuc are not secreted, transfected cells must be lysed, followed by intracellular substrate introduction and luminescence measurement. Furthermore, both FLuc and RLuc are large, making them less favorable protein tags.
由於FLuc及RLuc非分泌型，因此染的胞(引外源之DNA、RNA 或蛋白的胞)必裂解，然後行引入胞的酶作用物及光量。此外，FLuc及RLuc者皆大的，使得它是不佳的蛋白。
The secreted Gaussia luciferase (GLuc) is more stable. However, a relatively short luminescence half-life makes it unsuitable for many experimental settings. NLuc is an engineerd luciferase derived from deep sea luminescent shrimp.
分泌型Gaussia光素酶(GLuc)定。不，是一相上短的光半衰期，使其不合多境。NLuc是一，源自深海光，工程改造的光素酶。
This reporter gene has significantly attracted scientists for its greater stability, smaller size, and >150-fold increase in luminescence. Despite these benefits, they are not ideal for in vivo applications because its blue emission and their substrate (furimazine) has low solubility and bioavailability.
信息基因，由於其高的定性、小的尺寸及在光方面，增加超150倍。因此，一直大地吸引科家。管此些，不就活的用而言，它不理想。因，其出的光及其酶作用物(呋喃嗪)，具有低的溶解度及生物可使用性。
The major advantage of a bacterial bioluminescence system is that, since researchers have figured out the genetic pathways to synthesize all of the substrates required for the emission of light, no exogenous luciferin addition is needed.
由於，研究人已解，合成光所需之所有酶作用物的途。因此，菌生物光物系的主要是，需添加外源的光素。
However, its dependence on reduced riboflavin phosphate (FMNH2), which is available at a limited quantity in mammalian cells, and low intensity blue light emission makes it less favorable for eukaryotic systems. In addition, fungi possess a bioluminescent system with the synthetic genes elucidated, but the intensity of this system is still weak.
不，它依於哺乳物胞中，原之量有限的核素磷酸(FMNH2)，及出低度的光，使其不合真核胞物系。此外，真菌有一，具有明之合成基因的生物光物系。不，物系的度仍然很弱。
Luciferase reporter assays help determine whether a protein activates or suppresses transcription of the target gene. It also analyzes translation regulation by cis-elements of an mRNA present in the 5′-untranslated region (UTR) and 3′-UTR.
光素酶信息定有助於，定蛋白是否活化或抑制的基因的。它也藉由，存在於 5'-非(UTR)及3'-UTR中，mRNA的式元件(能因子合影附近基因表的短DNA片段)，分析。
2. 光素信息定，用光素酶光素的光反，作的基因子活性及基因表的一信息出。
Luciferin reporter assays employ a luciferase-luciferin light emitting reaction as a readout for target gene promoter activity and gene expression.
Scientists use recombinant DNA technology to develop promoter-reporter constructs. They fuse the regulatory region of the target gene with a luciferase reporter gene. A second DNA construct encodes proteins hypothesized to influence transcription. Researchers transfect a cell culture system such as HEK 293T cells with both constructs.
科家使用重的DNA技，子信息的成物。他使的基因的域一光素酶信息基因融合。第二DNA成物，被推影的蛋白。研究人以成物，染如HEK 293T胞(衍生自人胚胎胞的胞)等，胞培物系。
Typically, scientists examine luciferase expression two to three days after cell transfection. They lyse the transfected cells and place the contents in a reaction tube. Researchers add a suitable substrate to the reaction tube, such as luciferin, which enables luciferase to catalyze a chemical reaction that produces light.
通常，在胞染後，科家光素酶表，到三天。他裂解染的胞，此些容物置入反管中。研究人一合的酶作用物，如使光素酶能催化，生光之化反的光素，添加到反管中。
A luminometer can detect and quantify the amount of light produced in each reaction tube. The light intensity indicates the amount of target gene expression. If a protein upregulates target gene transcription, the cell expresses more luciferase molecules, which generates bright light. However, if the protein downregulates transcription, negligible luciferase expression occurs.
光度能及量化，於每一反管中，生的光量。此光度示的基因的表量。倘若蛋白上的基因，此胞使更多生明亮光的光素酶分子，作出表。不，倘若蛋白下，生可忽略的光素酶表。
Promoter analysis in bacteria or eukaryotic cells typically employs a single luciferase reporter assay. This assay requires an excess of luciferin and ATP. A single luciferase reporter assay follows the classical methodology where scientists transfect cells with a promoter-reporter construct, followed by lysis, substrate introduction, and detection. Besides gene expression analysis, this assay is suitable for high throughput screening of protein-protein interactions as well.
於菌或真核胞之胞中的子分析，通常用一光素酶信息定。定需要外的光素及三磷酸腺苷(ATP)。一光素酶信息定遵循古典方法，在此科家使用子信息的成物染胞，然後行裂解、引入酶作用物及。除了基因表分析之外，定也合，蛋白蛋白之交互作用的高理量。
Scientists designed the split luciferase complementation (SLC) method to study protein-protein interactions. This assay determines the effect of the physical contact between signaling proteins. The SLC method detects the factor that inhibits protein-protein interactions.
科家了，分裂光素酶互(SLC)法，研究蛋白蛋白的交互作用。此定定了，信蛋白之的物理接效果。SLC方法了，抑制蛋白蛋白之交互作用的因子。
It involves the genetic fusion of two inactive luciferase fragments with interacting proteins. The strongest luminescence corresponds to intense interactions between the protein pair and vice versa.
它涉及了，不活之光素酶片段交互作用的蛋白之因子融合。最的光於，在蛋白之的烈交互作用，且反之亦然。
Dual luciferase assays exploit the varying biochemical luminescence requirements of different species’ luciferase proteins. They allow the sequential quantitative measurement of FLuc and RLuc activities in a single protein extract.
重的光素酶定利用了，不同物之光素酶蛋白的不同生化光必需物。它允FLuc及RLuc活性，於一蛋白萃取物中的定量量。
Scientists use this method to simultaneously monitor different cellular events and their regulation based on two distinct signal emissions (e.g., green and red light). The improvement in DLR applicability relies on the availability of a luminometer equipped with two optical filters.
科家使用此方法同，不同的胞活，及其根不同信出(譬如光及光)的。此在重-光素酶之信息(DLR：Dual-Luciferase Reporter)用性的改善，依配光器之光度的可用性。
For example, researchers developed a DLR assay using two reporter genes (FLuc and RLuc) that quantifies sequential gene expression in yeast (Saccharomyces cerevisiae) using an advanced luminometer with wavelength-specific filters offering exceptional sensitivity for detecting distinct emissions.
譬如，研究人了一，使用信息基因(FLuc及RLuc)的DLR定方法。定方法使用一，具有提供不同光出特定波之光器的先光度，量化酵母菌(酒酵母)中，的基因表。
Multicolor luciferase reporter assays are powerful molecular biological tools that enable simultaneous monitoring of multiple genes with a single substrate. Scientists designed reporter genes for multicolor luciferase assays using three separate luciferase genes that emit green, orange, and red light upon addition of a single substrate.
多色的光素酶信息定是一，使以一酶作用物能同，多基因之有力的分子生物工具。科家使用三，在添加一酶作用物，出光、橙光及光之不同光素酶基因，了用於多色光素酶定的信息基因。
This method’s major advantage is the ability to monitor sequential and complex expression changes in multiple genes within a single cell or tissue. Researchers apply this assay for high-throughput gene expression analysis.
此方法的主要是，一胞或，於多基因中，且之表化的能耐。研究人此定法，用於高理量的基因表分析。
A luciferase-substrate reaction emits light ranging across many wavelengths. The sensitivity of in vivo bioluminescence imaging depends on light emission at the NIR range (700-900nm) and tissue permeability of the numberof photons emitted.
光素酶酶作用物的反出，跨多波的光。於活生物光造影的敏度，取於近外(NIR：Near Infrared)(700-900nm)出的光及出之光子量的穿透性。
Scientists developed a NIR luciferin analog to detect photons from deep target tissues with high sensitivity. Currently, many researchers are focusing on tuning the color of light emitted by bioluminating reporter genes towards the NIR or IR region, which has better tissue penetrance.
科家了一，似自深的之光子，具高敏度的近外光素。目前，多研究人正著重於，生物光信息基因出的光色，整到具有更佳穿透性的近外或外域。
Scientists are also working on multicomponent imaging to study several features or cell types simultaneously. The main concept behind this technique is the selective introduction of different luciferase molecules into separate cells within the same system. Upon specific substrate addition, only target cells light up.
科家也致力於多成分的造影，同研究多特徵或胞型。此技背後的主要概念是，不同的光素酶分子，性引入相同物系的不同胞。於添加特定酶作用物後，的胞光。
Scientists developed a multicolor bioluminescence system using NIR-emitting luciferases that enabled detection of two different cell populations within one animal model. A key shortcoming of this technique is the lack of a standardized method to visualize combinations of luciferase reporters.
科家使用出近外的光素酶了一，使能一物模型，不同胞群的多色生物光物系。此技的一主要缺是，缺乏化光素酶信息合的化方法。
址：https://www.the-scientist.com/luciferase-a-powerful-bioluminescent-research-tool-72013
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