4.5 Article

Molecular cloning, characterization and expression analysis of P53 from high latitude fish Phoxinus lagowskii and its response to hypoxia

Journal

FISH PHYSIOLOGY AND BIOCHEMISTRY
Volume 48, Issue 3, Pages 631-644

Publisher

SPRINGER
DOI: 10.1007/s10695-022-01072-6

Keywords

Phoxinus lagowskii; P53; Hypoxia; Cloning; Gene expression; FISH

Funding

  1. National Natural Science Foundation of China [32170523]
  2. Harbin Normal University Graduate Innovation Project [HSDSSCX2021-08]
  3. Outstanding Youth Science Fund Project of Harbin Normal University
  4. Blue granary scientific and technological innovation of China [2019YFD0900405]
  5. fundamental scientific research business expenses of colleges and universities in Heilongjiang Province [2021-KYYWF-0165]

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By cloning the cDNA of p53 from Phoxinus lagowskii, this study found that Pl-p53 is commonly expressed in various tissues and shows different expression patterns under different hypoxic conditions. These results provide a theoretical basis for understanding the changes of p53 in fish facing hypoxic environments.
As an intermediate link between multiple cellular stresses and cellular responses, p53, together with its upstream and downstream regulators and related genes, constitutes a complex network that regulates cellular stresses and cellular responses. However, no studies have investigated p53 in Phoxinus lagowskii, particularly the expression of p53 under different hypoxic conditions. In the present study, the cDNA of p53 from the Phoxinus lagowskii was cloned by the combination of homology cloning and rapid amplification of cDNA ends (RACE) approaches. The full-length cDNA of Pl-p53 was 1878 bp, including an open reading frame (ORF) of 1116 bp encoding a polypeptide of 371 amino acids with a predicted molecular weight of 41.22 kDa and a theoretical isoelectric point of 7.38. Quantitative realtime (qRT) PCR assays revealed that Pl-p53 was commonly expressed in all tissues examined, with highest expression in the heart. In addition, we investigated the expression of Pl-p53 in different tissues under different hypoxic conditions. In the short-term hypoxia group, Pl-p53 expression was down-regulated in both the brain and heart. The Pl-p53 expression was significantly elevated at 6 h in the muscle and liver, and was significantly up-regulated at 24 h in spleen. These results suggest that Pl-p53 plays different regulatory roles and provide a theoretical basis for the changes of p53 in fish facing hypoxic environments.

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