"Other Essentials"

Supplier: Bioss

Polyadenylation of mRNA precursors is a two-step reaction that requires multiple protein factors. The first step, endonucleolytic cleavage of polyadenylation substrates, requires CstF (cleavage stimulation factor), a heterotrimer that is composed of three distinct subunits. Heterotrimeric CstF recognizes GU- and U-rich sequences located downstream of the polyadenylation site on RNA. CstF-77 (cleavage stimulation factor, 77 kDa subunit), also known as CstF3, is one of the three subunits comprising CstF. It can exist as a homodimer and functions as the bridge, directly interacting with the other two CstF subunits, namely CstF-64 and CstF-50. CstF-77 is highly conserved among eukaryotes. It contains an Alpha-helical structure with 11 HAT (Half-a-TPR-containing) repeats and is essential for CstF assembly. In addition, CstF-77 is capable of interacting with CPSF1 and FCP1, other factors involved in polyadenylation.

Supplier: Bioss

BIVM (for basic, immunoglobulin like variable motif containing) refers to a recently identified gene product that maps to human chromosome 13q32-q33 and is predicted to encode a 503 amino acid protein. BIVM shows ubiquitous expression in normal human tissue and the presence of a 5' CpG island suggests it is a housekeeping gene. BIVM is likely essential for some aspect of basic cellular function. BIVM is highly charged and localizes to the cytoplasm and nucleus where it may bind to either DNA or RNA or associate with other cellular proteins. Significant sequence homology exists with many organisms.

Supplier: ProSci Inc.

Interleukin-2(IL-2) is an interleukin, a type of cytokine signaling molecule in the immune system，belongs to the IL-2 family. It is a powerful immunoregulatory lymphokine produced by T-cells in response to antigenic or mitogenic stimulation. IL-2/IL-2R signaling is required for T-cell proliferation and other fundamental functions that are essential for the immune response. IL-2 stimulates growth and differentiation of B-cells, NK cells, lymphokine-activated killer cells, monocytes, macrophages and oligodendrocytes.

Supplier: ProSci Inc.

Ubiquitin-Conjugating Enzyme E2 B (UBE2B) belongs to the ubiquitin-conjugating enzyme family. UBE2B can catalyze the covalent attachment of ubiquitin to other proteins, and is essential for the multi-ubiquitination and degradation of N-end rule substrates. UBE2B is indispensability for postreplication repair of UV-damaged DNA and may be involved in neurite outgrowth.Additionally, UBE2B may have a role in sepsis-induced muscle protein proteolysis and cancer-induced cachexia.

Supplier: G-Biosciences

Concanavalin A (Con A) agarose consists of Con A coupled to 6% agarose by the cyanogen bromide method. Con A is a tetrameric metalloprotein lectin isolated from Canavalia ensiformis (jack bean). It is used for the purification of glycoproteins, polysaccharides and glycolipids as it binds molecules containing α-D-mannopyranosyl, α-D-glucopyranosyl and sterically related residues. It has also be used in other application areas including purification of enzyme-antibody conjugates, purification of IgM and separation of membrane vesicles. As stated above, it is a metalloprotein and to maintain its binding characteristics the presence of both Mn²⁺ and Ca²⁺ is essential. Each subunit of Con A utilises one calcium and one manganese ion and these cations can be removed under acidic conditions abolishing the carbohydrate-binding activity.

Supplier: Bioss

RNA exonuclease that binds to the 3'-end of histone mRNAs and degrades them, suggesting that it plays an essential role in histone mRNA decay after replication. A 2' and 3'-hydroxyl groups at the last nucleotide of the histone 3'-end is required for efficient degradation of RNA substrates. Also able to degrade the 3'-overhangs of short interfering RNAs (siRNAs) in vitro, suggesting a possible role as regulator of RNA interference (RNAi). Requires for binding the 5'-ACCCA-3' sequence present in stem-loop structure. Able to bind other mRNAs. Required for 5,8S rRNA 3'-end processing. Also binds to 5,8S ribosomal RNA. Binds with high affinity to the stem-loop structure of replication-dependent histone pre-mRNAs.

Supplier: Bioss

RNA exonuclease that binds to the 3'-end of histone mRNAs and degrades them, suggesting that it plays an essential role in histone mRNA decay after replication. A 2' and 3'-hydroxyl groups at the last nucleotide of the histone 3'-end is required for efficient degradation of RNA substrates. Also able to degrade the 3'-overhangs of short interfering RNAs (siRNAs) in vitro, suggesting a possible role as regulator of RNA interference (RNAi). Requires for binding the 5'-ACCCA-3' sequence present in stem-loop structure. Able to bind other mRNAs. Required for 5,8S rRNA 3'-end processing. Also binds to 5,8S ribosomal RNA. Binds with high affinity to the stem-loop structure of replication-dependent histone pre-mRNAs.

Supplier: Bioss

RNA exonuclease that binds to the 3'-end of histone mRNAs and degrades them, suggesting that it plays an essential role in histone mRNA decay after replication. A 2' and 3'-hydroxyl groups at the last nucleotide of the histone 3'-end is required for efficient degradation of RNA substrates. Also able to degrade the 3'-overhangs of short interfering RNAs (siRNAs) in vitro, suggesting a possible role as regulator of RNA interference (RNAi). Requires for binding the 5'-ACCCA-3' sequence present in stem-loop structure. Able to bind other mRNAs. Required for 5,8S rRNA 3'-end processing. Also binds to 5,8S ribosomal RNA. Binds with high affinity to the stem-loop structure of replication-dependent histone pre-mRNAs.

Supplier: Bioss

RNA exonuclease that binds to the 3'-end of histone mRNAs and degrades them, suggesting that it plays an essential role in histone mRNA decay after replication. A 2' and 3'-hydroxyl groups at the last nucleotide of the histone 3'-end is required for efficient degradation of RNA substrates. Also able to degrade the 3'-overhangs of short interfering RNAs (siRNAs) in vitro, suggesting a possible role as regulator of RNA interference (RNAi). Requires for binding the 5'-ACCCA-3' sequence present in stem-loop structure. Able to bind other mRNAs. Required for 5,8S rRNA 3'-end processing. Also binds to 5,8S ribosomal RNA. Binds with high affinity to the stem-loop structure of replication-dependent histone pre-mRNAs.

Supplier: ProSci Inc.

CDC23 shares strong similarity with Saccharomyces cerevisiae Cdc23, a protein essential for cell cycle progression through the G2/M transition. This protein is a component of anaphase-promoting complex (APC), which is composed of eight protein subunits and highly conserved in eukaryotic cells. APC catalyzes the formation of cyclin B-ubiquitin conjugate that is responsible for the ubiquitin-mediated proteolysis of B-type cyclins. This protein and 3 other members of the APC complex contain the TPR (tetratricopeptide repeat), a protein domain important for protein-protein interaction.The protein encoded by this gene shares strong similarity with Saccharomyces cerevisiae Cdc23, a protein essential for cell cycle progression through the G2/M transition. This protein is a component of anaphase-promoting complex (APC), which is composed of eight protein subunits and highly conserved in eucaryotic cells. APC catalyzes the formation of cyclin B-ubiquitin conjugate that is responsible for the ubiquitin-mediated proteolysis of B-type cyclins. This protein and 3 other members of the APC complex contain the TPR (tetratricopeptide repeat), a protein domain important for protein-protein interaction.

Supplier: ProSci Inc.

Human proteins MECP2, MBD1, MBD2, MBD3, and MBD4 comprise a family of nuclear proteins related by the presence in each of a methyl-CpG binding domain (MBD). Each of these proteins, with the exception of MBD3, is capable of binding specifically to methylated DNA. MECP2, MBD1 and MBD2 can also repress transcription from methylated gene promoters. In contrast to other MBD family members, MECP2 is X-linked and subject to X inactivation. MECP2 is dispensible in stem cells, but is essential for embryonic development. MECP2 gene mutations are the cause of some cases of Rett syndrome, a progressive neurologic developmental disorder and one of the most common causes of mental retardation in females.DNA methylation is the major modification of eukaryotic genomes and plays an essential role in mammalian development. Human proteins MECP2, MBD1, MBD2, MBD3, and MBD4 comprise a family of nuclear proteins related by the presence in each of a methyl-CpG binding domain (MBD). Each of these proteins, with the exception of MBD3, is capable of binding specifically to methylated DNA. MECP2, MBD1 and MBD2 can also repress transcription from methylated gene promoters. In contrast to other MBD family members, MECP2 is X-linked and subject to X inactivation. MECP2 is dispensible in stem cells, but is essential for embryonic development. MECP2 gene mutations are the cause of some cases of Rett syndrome, a progressive neurologic developmental disorder and one of the most common causes of mental retardation in females.

Supplier: Bioss

The protease enzyme Lethal Factor (LF) is one of the three proteins (LF, EF & PA) composing the anthrax toxin produced by Bacillus anthracis, a bacteria which can infect many mammalian species and that may be fatal. LF is not toxic by itself, but when associated with Protective Antigen (PA), can then gain entry to cells. Once inside the cell, LF then cleaves the N terminal of most dual specificity mitogen activated protein kinase kinases (MAPKKs or MAP2Ks) (except for MAP2K5). Cleavage invariably occurs within the N terminal proline rich region preceding the kinase domain, thus disrupting a sequence involved in directing specific protein protein interactions necessary for the assembly of signaling complexes. There may be other cytosolic targets of LF involved in cytotoxicity. The proteasome may mediate a toxic process initiated by LF in the cell cytosol involving degradation of unidentified molecules that are essential for macrophage homeostasis. This is an early step in LF intoxication, but it is downstream of the cleavage by LF of MEK1 or other putative substrates.

Supplier: ProSci Inc.

MCM8 is one of the highly conserved mini-chromosome maintenance proteins (MCM) that are essential for the initiation of eukaryotic genome replication. The hexameric protein complex formed by the MCM proteins is a key component of the pre-replication complex (pre_RC) and may be involved in the formation of replication forks and in the recruitment of other DNA replication related proteins. This protein contains the central domain that is conserved among the MCM proteins. This protein has been shown to co-immunoprecipitate with MCM4, 6 and 7, which suggests that it may interact with other MCM proteins and play a role in DNA replication. Alternatively spliced transcript variants encoding distinct isoforms have been described.

Supplier: Bioss

RNA exonuclease that binds to the 3'-end of histone mRNAs and degrades them, suggesting that it plays an essential role in histone mRNA decay after replication. A 2' and 3'-hydroxyl groups at the last nucleotide of the histone 3'-end is required for efficient degradation of RNA substrates. Also able to degrade the 3'-overhangs of short interfering RNAs (siRNAs) in vitro, suggesting a possible role as regulator of RNA interference (RNAi). Requires for binding the 5'-ACCCA-3' sequence present in stem-loop structure. Able to bind other mRNAs. Required for 5,8S rRNA 3'-end processing. Also binds to 5,8S ribosomal RNA. Binds with high affinity to the stem-loop structure of replication-dependent histone pre-mRNAs.

Supplier: Bioss

Mapping to chromosome 21, the FASP1 gene (FAPP1-associated protein 1) encodes a 233 amino acid protein that is homologous to the fission yeast protein Mis18. In yeast, Mis18 is localized to the centrosome and forms a complex with Mis16 to maintain the deacetylated state of histones specifically in the central core of centromeres. FASP1, also known as Protein Mis18-alpha and C21orf45, is required for the recruitment of CENP-A to centrosomes and is thereby essential for normal chromosome segregation during mitosis. With expression in testis, FASP1 exists as a homodimer, a heterodimer with MIS18B or is present in a complex containing other Mis18 family members. FASP1 has been shown to bind to pp5644 in Hela cells, where overexpression of pp5644 leads to inhibited growth and colony formation.

Supplier: G-Biosciences

The final goal in molecular biology is often the expression of a recombinant protein. The transformed plasmids can be used as templates by the bacteria to produce protein. Students learn about essential promoters and other elements necessary for successful protein expression in bacteria, including the differences between inducible and constitutive (unregulated) expression.The Expression of a Recombinant Protein kit allows students to express a protein either constitutively or under the control of an inducible promoter, which is activated with IPTG (isopropyl-beta-D-thiogalactopyranoside).