Cloning, Expression, And Genetical Transformation Analysis Of A Calmodulin-like Gene Which Is Induced By Multiple Stresses In Rice(Oryza Sativa L.)

To discover and identify new stress-tolerance genes and improve crop stress tolerance to obtain steady and high yields of crop productions, we performed microarray(genechip)analysis of rice Peiai64s(Oryza sativa L.)using the GeneChip(?)Rice Genome Array(Affymetrix) representing 51,279 transcripts from two rice cultivars japonica and indica..Results showed that activities of many genes were evidently increased or decreased.OsMsr-2 is one of the genes that could be induced by cold,drought and heat.The data obtained in this research mainly include the followings:Firstly,we found that OsMsr-2 could be induced by clod,drought and heat by microarray analysis.The data showed that the expression level of OsMsr-2 was elevated in different stages and different tissues, with the average fold of 19.9 increased,ranging from 3.5 to 68.2 folds.Secondly,we performed real-time RT-PCR analysis to further validate the expression pattern of OsMsr-2.The results confirmed that OsMsr-2 indeed could be induced by multiple stresses,similar to the result obtained by the microarray analysis.Thirdly,we performed bioinformatics analysis and found that OsMsr-2 has two EF-Hand domains,which is similar to a calmodulin-like gene.Analyzing the promoter region identified some cis-acting elements related to stress responses.The alignment of peptide sequences revealed that OsMsr-2 is closely related to the calmodulin-like sub-family which might play an important role in the signal transduction in rice.Fourthly,we designed specific primers according to homologous gene sequences found in Genbank and cloned the cDNA sequence containing the whole open reading frame of the gene.Fifthly,we constructed gene over-expression vector and delivered the gene into Arabidopsis successfully.The analysis of transgenic Arabidopsis is in progress.