|Table of Contents|

Screening of SIVA-1 gene interacting proteins in drug-resistant human gastric cancer cells by yeast two hybrid

Journal Of Modern Oncology[ISSN:1672-4992/CN:61-1415/R]

Issue:
2023 05
Page:
806-811
Research Field:
Publishing date:

Info

Title:
Screening of SIVA-1 gene interacting proteins in drug-resistant human gastric cancer cells by yeast two hybrid
Author(s):
YANG Jie1; LI Wei1; LIU He1; KONG Fanbiao2; WANG Xiaotong3
Keywords:
gastric cancer; SIVA-1 gene; yeast two-hybrid; PCBP1
PACS:
R735.2
DOI:
10.3969/j.issn.1672-4992.2023.05.004
Abstract:
Objective：To screen the proteins interacting with SIVA-1 gene in drug-resistant human gastric cancer cells by yeast two hybrid technique.Methods:The cDNA library of human gastric cancer drug-resistant cells was constructed,and the related proteins of SIVA-1 were screened by yeast two hybrid and in vitro immunoprecipitation assay,so as to explore the mechanism of its effect on the drug resistance of gastric cancer drug-resistant cells.Results:The cDNA library of drug-resistant human gastric cancer cells was successfully constructed,and the titer was 4.78×106 cfu/mL,recombination rate>95%,and the titer was 9.50×109 pfu/mL after amplification.The bait expression vector pGBKT7-SIVA-1 was successfully constructed,and the self activation test showed that the recombinant bait vector pGBKT7-SIVA-1 did not have self-excited activity.After preliminary screening,we obtained 21 positive clones that can activate three reporter genes (Ade2,His3 and LacZ) at the same time.The 21 positive clones were further extracted for plasmid,amplified and yeast rotation experiment was performed.Finally,four positive candidate clones interacting with SIVA-1 truncate were obtained.After gene sequencing of the four library inserts,two different proteins interacting with SIVA-1 were obtained:UXT (ubiquitin transcript) and PCBP1 (polycytosine binding protein 1).After in vitro immunoprecipitation experiment,PCBP1 interacting with gastric cancer drug-resistant cell SIVA-1 was finally screened.Conclusion:There is a protein-protein interaction between PCBP1 and SIVA-1.

References:

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Memo

Memo:
National Natural Science Foundation of China（No.81660416，No.82260468）;国家自然科学基金项目（编号：81660416，82260468）
Last Update: 2023-01-31