Cloning And Sequencing Analysis Of Heme Oxygenase-1(HO-1) Gene From Ochotona Curzoniae

Background and ObjectivesOchotona curzoniae is small mammals of qinghai-tibet plateau which is Lagomorpha,Ochotonidae and Ochotona. Ochotona curzoniae belongs to generation living in3200~4700meters above sea level of the hypoxia, extremely cold alpine meadow andsteppe. It has high efficiency mechanism in the process of energy metabolism andoxygen transfer.Therefore, Ochotona curzoniae can tolerate hypoxia and lowtemperature. Making it an important constituent of plateau ecological system and anideal animal of hypoxic adaptation research.Studies have found that many factors can cause the expression of induction type HO(HO-1) increase. HO-1and hemoglobin degradation products biliverdin, bilirubin,carbon monoxide and iron ions are involved in many physiological and pathologicalprocess which play a very important adjustment. HO-1protects cells by inhibiting theproduction of inflammatory cytokines, acting anti-oxidation stress, attenuatinginflammation response in oxidative stress state.This study provides the plateau hypoxia adaptation experimental data from gene levelby cloning and analyzing the heme oxygenase-1(HO-1) gene from Ochotonacurzoniae.MethodsOchotona curzoniaes were captured in muli town tianjun county Qinghaiprocince(about4062m above sea level) in July2011.The average weight was200g.Physiological indicators were collected and liver was extrscted and stored in liquidnitrogen after intraperitoneal injection of25%urethane anesthesia. Amplificationprimers were designed by Primer5.0and DNAMAN6.0according the relativelyconservative area of HO-1cDNA sequence of cows, mice, rats and othermammals species published in GenBank. The HO-1cDNA was cloned by RT-PCR and RACE from an Ochotona curzoniae’s liver. The bioinformatics of HO-1genewere then performed.ResultsThe cDNA of HO-1gene of Ochotona curzoniae was obtained and deposited in GenBank asaccession number JX035934. The full length of cDNA was1466bp, including873bp encoding sequence (290amino acids). Homology comparison showed that the DNA sequence of the HO-1gene was highly homologousand the homology more then80%with Oryctolags cuniculus, Homo sapiens,Bos Taurus, Mus musculus, Rattus norvegicus, Sus scrofa, and Equus caballus.In this study,5（4th，92th，223th，250th，255th） loci of15amino aciddifferences sites may lead to regulate interactions betweenHO-1and hypoxiafactor.ConclusionThe cDNA of HO-1gene of Ochotona curzoniae was obtained and deposited first inGenBank as accession number JX035934.5（4th，92th，223th，250th，255th） lociof15amino acid differences sites may lead to regulate interactions betweenHO-1and hypoxia factor. Compared with the same moving plateau species, multiple locimutations were occurred. It showed that localized plateau pika have significantdifferences on the structure and function, this may be the expression of geneticadaptation in plateau pika, also may be one of the reasons for its strong hypoxiaresistance, but the specific mechanism of mutations associated with plateau geneticadaptation have remained to be further research.HO-1cytoprotective closely related to the above findings for a better understanding ofthe plateau pika protect cells from oxidative stress injury mechanism, by further adaptof altitude hypoxia alpine gene plateau pika for the futuregood early intervention theoxidative damage progression of the disease, cure oxidative damage caused by thedisease of major significance.