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Analysis of JAK2 Gene Rearrangement by Southern Blot Technology (CAT#: STEM-MHT-0021-LGZ)

Introduction

Official Full Name: Janus kinase 2<br />Also known as: JTK10<br />This gene encodes a non-receptor tyrosine kinase that plays a central role in cytokine and growth factor signaling. The primary isoform of this protein has an n-terminal FERM domain required for erythropoietin receptor binding, an SH2 domain that binds STAT transcription factors, a pseudokinase domain, and a c-terminal tyrosine kinase domain. Cytokine binding induces autophosphorylation and activation of this kinase. The kinase then recruits and phosphorylates signal transducers and activators of transcription (STAT) proteins. Growth factors like TGF-β1 also induce phosphorylation and activation of this kinase and translocate downstream STAT proteins to the nucleus, where they affect gene transcription. Mutations in this gene are associated with many inflammatory diseases and malignancies. This gene is a downstream target of the pleiotropic cytokine IL6 produced by B cells, T cells, dendritic cells, and macrophages, generating an immune response or inflammation. Dysregulation of the IL6/JAK2/STAT3 signaling pathway can lead to increased proliferation of hematopoietic stem cells and myeloproliferative neoplasms. Nonsynonymous mutations in the pseudokinase domain of this gene disrupt the repressive role of this domain, resulting in constitutive tyrosine phosphorylation activity and hypersensitivity to cytokine signaling. This gene and the IL6/JAK2/STAT3 signaling pathway are therapeutic targets for the treatment of excessive inflammatory responses to viral infections. Alternative splicing results in multiple transcript variants encoding different isoforms.




Principle

Under certain conditions, two single strands of nucleic acid with certain homology can be specifically hybridized to form double strands according to the principle of base complementarity. Generally, DNA molecules to be detected are digested with restriction enzymes, separated by agar-gel electrophoresis, denatured and transferred to nitrocellulocellulose film or nylon film or other solid phase support according to their position in the gel, fixed and then reacted with DNA probes labeled with isotopes or other markers. This is followed by autoradiography or an enzyme reaction to detect the amount of specific DNA molecules. If the object to be tested contains a sequence that is complementary to the probe, the two are combined by the principle of base complementarity, and the free probe is washed and detected by self-development or other suitable techniques, thus revealing the fragment to be tested and its relative size.

Applications

Gene Rearrangement Detection

Procedure

1. Sample Processing
2. DNA Extraction and Digestion
3. Gel Electrophoresis
4. Gel Pretreatment
5. Transfer membrane
6. Probe Labeling
7. Prehybridization (blocking)
8. Southern hybridization
9. Membrane washing
10. Autoradiographic Assay
11. Results Analysis

Materials

Sample: DNA, Bacterial Fluid/Tissue/Cell