Cloning And Functional Analysis Of Gene OsHAD1 In Rice

Rice is an important cash crop,and one third of the world's population is rice-based.At present,rice has been genome-wide sequencing,so it is often studied as an important model plant.In agricultural production,various abiotic stresses such as high salt,drought,chilling injury become an important factor affecting food production.Therefore,it is very important to study the resistance mechanism of rice and the improvement of rice yield.OsSCL30b is a rice gene selected by this laboratory.The gene encoded by the gene belongs to the RRM family.RNA recognition motif(RRM),also known as ribonucleoprotein domain(RNP)or RNA binding domain(RBD).The protein domain is indispensable for the binding of RNA molecules to specific substances and domains.RNA recognition motif is a protein domain that is widely present in eukaryotes.The eukaryotic RRM protein is present in all post-transcriptional events:pre-mRNA processing,alternative cleavage,mRNA stability,RNA editing,mRNA export,pre-rRNA complex formation and degradation.RRM proteins are present in the chloroplasts of plants and are involved in the 3 'end of chloroplast mRNA processing.The growth of OsSCL30b transgenic plants in the field showed a later than the growth period of Japanese sunny.In order to study the mechanism of OsSCL30b transgenic plants in OsSCL30b,OsSCL30b gene was used as bait protein,and some proteins interacting with OsSCL30b were screened from rice cDNA library by yeast two-hybrid system.A protein named OsHAD1 to carry out research.The HAD superfamily is named after the halogenated acid dehalogenase,including various hydrolases,including phosphatase,phosphorylasc and phosphogluconasc.The members of the family of proteins are involved in the metabolism of glycerol metabolism and glycolytic phosphate pathway in Arabidopsis thaliana.Bioinformatics analysis of the gene showed that there were many stress-related elements in the promoter region of the gene,suggesting that there may be some resistance to stress in stress physiology.Based on this,this study mainly explored the expression of the gene under adversity.And to provide a basis for studying the role of OsSCL30b and the relationship between rice growth delay and stress physiology.The main results are as follows:The OsHAD1 gene encodes 283 amino acids with an isoelectric point of 5.23.The cis-acting elements in the upstream promoter are ABRE,LTR,MBS,TGACGand CGTCA,TCA-element and so on.In this study,the cDNA was induced by RT-PCR,and the cDNA was induced by different stress.It was especially obvious under low temperature and drought.The prokaryotic expression vector of OsHAD1 gene was constructed and expressed in vitro.The growth status of Escherichia coli expressing OsHAD1 protein under adverse conditions was also investigated.The results showed that the expression of Escherichia coli could increase the expression of E.coli Low temperature tolerance,reducing the tolerance of E.coli to drought and salt.Construction of OsHAD1 gene overexpression vector,OsHAD1 gene expression vector was constructed and transformed into rice callus by Agrobacterium tumefaciens infection,and transgenic plants were obtained by differentiation.And detected by hygrormycin immersion and hygromycin primer and 35S primer for the correct positive transgenic plants.Which provided the material for the further study of OsHAD1 protein in eukaryotic rice.