Application of VGLL4 gene in treatment of tumors and related medicament of VGLL4 gene
A gene and drug technology, applied in the biological field, can solve problems such as excessive cell proliferation and increased risk of cancer
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[0091] Example 1 The relationship between the expression of VGLL4 and the occurrence and development of gastric cancer
[0092] 1. Experimental method
[0093] In order to study the clinical relationship between VGLL4 and YAP, the mRNA levels of VGLL4 and YAP in 84 human gastric cancer samples and 91 normal human gastric tissue samples were detected. Guanidine isothiocyanate is used to extract total RNA in one step. AMV reverse transcriptase synthesizes the first strand and uses it as a template to perform PCR amplification with corresponding primers under the action of DNA taq enzyme; PCR primers are shown in Table 1, Realtime -The PCR reaction system is shown in Table 2, the Realtime-PCR amplification program is shown in Table 3, and the PCR amplification results are shown in figure 1 A and figure 1 B.
[0094] Table 1 Primer sequence
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[0096] Table 2 Realtime-PCR reaction system
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[0098] Table 3 PCR reaction system program setting
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[0100] A tumor sample o...
Example Embodiment
[0106] Example 2 VGLL4 inhibits YAP-mediated TEAD4 transcriptional activation and cell proliferation
[0107] 1. The purpose of the experiment
[0108] The purpose of this example is to study the regulation modes of VGLL4, YAP and TEAD4 and their influence on cell proliferation.
[0109] 2. Experimental method
[0110] 2.1 Plasmid construction
[0111] 1) Construction of pCDH-YAP plasmid and pCDH-VGLL4 plasmid: One-step extraction of total RNA from human HEK293 cells with guanidine isothiocyanate, AMV reverse transcriptase to synthesize the first strand, and use it as a template for high-fidelity DNA polymerization in Takara Enzyme PrimeSTAR Use the corresponding primers to perform PCR amplification under the action of, the PCR primers are shown in Table 4, the PCR reaction system is shown in Table 5, and the PCR amplification program is shown in Table 6.
[0112] Table 4 Primer sequence
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[0114] Table 5 PCR reaction system
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[0116] Table 6 PCR amplification program
[0...
Example Embodiment
[0137] Example 3 Vgl4-TEAD4 interaction affects Vgl4 inhibits YAP-mediated transcriptional activation of TEAD4 and its downstream genes
[0138] 1. Experimental materials
[0139] 1.1 Construction of plasmid
[0140] 1) HT-pET-28a vector: Take commercial pET-28a (Novagen) as a template and use the following primers for construction. The PCR amplification program is: 98°C 2min; PCR cycle: 98°C 10s, 58°C 10s, 68°C 1kb / min, 18 cycles; 68°C 10min. Use DpnI demethylase to remove the unmutated wild-type pET28a template, transform the PCR amplified fragment into DH5α competent cells, and use the characteristic of repairing vector deletion in E. coli itself to circularize the entire fragment to form a vector through resistance After screening, the obtained HT-pET-28a vector with the same backbone as the original pET28a vector, but with TEV restriction sites and commonly used multiple cloning sites.
[0141] HT-s:
[0142] 5’-Gaaaacctgtattttcagggcgccatggatccggaattcaaaggcctacgtcgacgagctcaactag...
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