Screening,Cloning And Bioinformatics Analysis Of Genes Related To Drought Environment Adaptation In Tarim Red Deer

Tarim red deer(Cervus elaphus yarkanensis)which lives in the harsh desert environment of Tarim Basin,plays an important role in the desert community of in China,and is the only special subspecies that highly adapts to the desert environment,so it has unique research and development value.In recent years,although there have been many reports on the population number,genetic diversity and genetic structure of Tarim red deer,but the molecular mechanism of Tarim red deer adapting to drought environment is still unclear.In this study the genes related to adaptation of Tarim red deer to arid environment were screened on the bases of genome resequencing and transcriptome sequencing data of Tarim red deer then were cloned,sequenced and bioinformatics analyzed.The main results are as follows:(1)By using the method of F_ST&?and XP-EHH selective elimination analysis and consulting a large number of relevant literatures,we have screened the PRDX3 gene from whole genome resequenced data of Tarim red deer in our research group with F_ST value of 0.791,?value of 1.075 and XP-EHH value of 2.869.This gene was significantly enriched in some important biological processes and plays most important biological functions,such as RNA metabolism process(GO:0016070),transcription process(GO:0006351),protein secretion(GO:0005634),ion binding(GO:0043167),molecular function(GO:0003674)and oxidoreductase activity(GO:0016628),meanwhile the gene was enriched in KEGG metabolic pathway(bta:01100).(2)Based on the transcriptome sequencing data of our research group,and the combined with consulting and referring to relevant literatures,we have screened the AQP1 gene which was enriched into glomerular filtration function(GO:0003094),renal water absorption(GO:0070295),water channel activity(GO:0015250),as well as enriched in bile secretion(ko04976)and proximal tubule bicarbonate reclamation(ko04964)pathways.(3)The m RNA expression of PRDX3 and AQP1 genes were analyzed by q RT-PCR.The results showed that,PRDX3 and AQP1 genes had relatively high m RNA expression level in the kidney tissue of Tarim red deer.(4)The PRDX3 and AQP1 genes of Tarim red deer were amplified by RT-PCR.Then the amplified samples were cloned and sequenced,and the sequencing sequence were proofread,sorted and spliced manually.Finally,the CDS sequences of PRDX3 and AQP1genes of Tarim red deer,were obtained with the length of 660 bp and 360 bp,respectively.The results were consistent with the expected results.(5)The bioinformatics technology was used to analyze the CDS region of PRDX3gene of Tarim red deer.The results showed that,PRDX3 gene was relatively conservative.PRDX3 is composed of 220 amino acids.PRDX3 protein with instability coefficient of25.36,molecular weight of 24.42 ku,total average hydrophilicity of-0.05,and fat solubility coefficient of 85.50 showed fat soluble and relatively stable hydrophilic protein.There were abundant phosphorylation sites and glycosylation sites in PRDX3 protein.The structure of this protein is relatively loose,and contains the conserved domain of PRX?typ2?cys superfamily and has relatively strong interactions with a variety of proteins.(6)According to the bioinformatics analysis of the CDS region of AQP1 gene in Tarim red deer,the AQP1 gene was relatively conservative.AQP1 is composed of 120amino acids.The molecular weight of AQP1 protein was 12.63 ku,the instability coefficient was 24.62,total average hydrophilicity was 0.91,and fat lipolysis coefficient was 122.00.It was a relatively stable hydrophobic protein with lipid solubility.AQP1 was a transmembrane protein withabundant phosphorylation and glycosylation sites.AQP1,which contains conserved domain of MIP superfamily and has a relatively strong interaction with many proteins.In this study,the genes related to adaptation of Tarim red deer to drought environment were studied at the molecular level.It would be foundation for the further study of functional genes in the Tarim red deer and provide valuable experimental evidence.