Cloning and Identification and Research on Radiation-resistant Ability of the HdNINJ1 Gene of Hypsibius dujardini
Tardigrades,known as the"water bears",are kinds of small invertebrates that can tolerate ultra-strong radiation.However the mechanism of radiation tolerance remains largely unknown.The aim of this study was to identify the potential radiation resistant genes and provide new insights into the study of radiation tolerance in tardigrades.From Hypsibius dujardini,we found a new gene ho-mologous to human nerve injury induced protein(Ninjurin)which was named HdNINJ1.In this study,using the cDNA of Hypsibius dujardini as a template,the coding sequence(CDS)of HdNINJ1 gene were obtained by PCR.The sequence characteristics were analyzed using bioinformatics methods,and the anti-radiation function of HdNINJ1 was detected through colony formation,alkaline comet and immunofluorescence assay.The results showed that the CDS of the HdNINJ1 gene had a total length of 675 bp,encoding 224 amino acids,and containing a Ninjurin domain.The results of colony formation assays showed that the survival rate of human U2OS cells expressing HdNINJI gene increased by 50.9％compared to the control group after 4 Gy X-ray irradiation.Comet assay and immunofluorescence assay results showed that human U2OS cells expressing HdNINJ1 gene were reduced in DNA damage by 21.0％and 22.9％after being irradiated with 10 Gy and 2 Gy X-rays,respectively.The results were statistically significant.This study indi-cates that HdNINJ1 can promote DNA damage repair in human U2OS cells and improve their radiation tolerance.
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