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Analysis of VHLL Gene (Mutation) by RT-qPCR (CAT#: STEM-MT-0760-LGZ)

Introduction

Official Full Name: VHL like<br />Also known as: VLP; VHLP<br />Von Hippel-Lindau (VHL) tumor suppressor protein is a component of an E3 ubiquitin ligase complex that selectively ubiquitinates the alpha subunit of the hypoxia-inducible factor (HIF) transcription factor for proteasome-mediated degradation. VHL inactivation leads to VHL disease and sporadic renal carcinoma. The VHL homologue encoded by this gene lacks one of two key domains required for VHL function. This gene may be involved in the regulation of oxygen homeostasis and neovascularization during placental development. The gene lacks introns and can also be interpreted as a reverse-transcribed pseudogene located at the VHL locus on chromosome 3. However, this protein is represented in this RefSeq due to evidence in PMID:14757845 that strongly suggests that the protein is translated. The same article also states that this protein binds HIFα but cannot recruit the E3 ubiquitin ligase complex, so it acts as a dominant-negative VHL protein and protector of HIFα.




Principle

Quantitative reverse transcription PCR (RT-qPCR) is an experimental method applied to PCR experiments using RNA as the starting material. In this method, total or messenger RNA (mRNA) is first transcribed into complementary DNA (cDNA) by reverse transcriptase. Subsequently, qPCR reaction was performed using cDNA as template.

Applications

Gene mutation analysis.

Procedure

1. Sample processing and preparation of PCR reaction system.
2. Add the amplification template, cover the PCR reaction cover, mix well, centrifuge at low speed instantaneously, and transfer to the PCR instrument.
3. Set the program for PCR amplification.
4. Data analysis.

Materials

Sample: depends on the customer's analysis requirements