What is fluorescent fusion?
The simplest fluorescent fusion proteins (FFPs) are FPs fused to targeting sequences (e.g., a nuclear localization sequence or a signal sequence) for the highlighting of an organelle or a cellular domain of interest.
What is GFP fusion protein?
INTRODUCTIONGFP (green fluorescent protein) fusion proteins have been used to address a wide range of questions in individual cells, as well as in tissues of a particular organism. GFP fusion proteins can be transiently or stably expressed.
How does GFP tagging work?
GFP-tagging is a way of preparing a sample for fluorescence microscopy by using the GFP as a fluorescent protein reporter. This is done by cloning the GFP in frame with the target protein at either the N- or C-terminus of the amino acid chain.
What is pEGFP N1?
pEGFP-N1 encodes a red-shifted variant of wild-type GFP (1–3) which has been optimized for brighter fluorescence and higher expression in mammalian cells. (Excitation maximum = 488 nm; emission maximum = 507 nm.)
What is the size of GFP?
GFP is a 28 kDa protein that resembles a cylinder with a length of 4.2 nm and a diameter of about 2.4 nm (Hink et al., 2000). The complete beta-barrel is necessary for its fluorescence and therefore GFP cannot be downsized by deleting residues.
What causes GFP fluorescence?
1. GFP is a barrel shape with the fluorescent portion (the chromophore) made up of just three amino acids. When this chromophore absorbs blue light, it emits green fluorescence.
How are fusion proteins expressed?
Fusion proteins may be expressed in cell-based and cell-free systems.
How are fusion proteins made?
A protein made from a fusion gene, which is created by joining parts of two different genes. Fusion genes may occur naturally in the body by transfer of DNA between chromosomes. For example, the BCR-ABL gene found in some types of leukemia is a fusion gene that makes the BCR-ABL fusion protein.
What is fusion tag?
A fusion tag is a known protein or peptide that is fused onto your protein of interest. As these tags are well characterized there is a wide range of top-performing antibodies available, enabling easy detection of a specific protein for a variety of applications.
What is in-fusion cloning technology?
A typical use for this technology would be to clone PCR products into vectors, without the use of restriction enzymes, ligase or phosphatase. In- Fusion cloning kits, which contain our proprietary In-Fusion Enzyme, let you rapidly generate very precise constructs.
What is in-fusion seamless cloning protocol?
The In-Fusion cloning protocol. Clone any insert, with any vector, at any site. In-Fusion seamless cloning technology makes it easy! Here’s how.
What is the in-fusion PCR method?
The In-Fusion method is simple and efficient. First, PCR primers are designed that share 15 bases of homology with the sequence at the ends of the linearized cloning vector (i.e., at the desired site of insertion; refer to Section V of this manual). These primers are then used to PCR amplify the insert DNA.
What is the in-fusion HD cloning system?
The In-Fusion HD Cloning System has an improved capability for cloning multiple fragments in a single reaction. Using this system, cloning up to four 1-kb fragments simultaneously is as easy as cloning a single fragment. This saves weeks that would otherwise be spent screening clones and subcloning.