Molecular Cloning And Analysis Of Histone H3 Genes From Triticum Aestivum

In order to further undersranding the functions of histon H3,we hoped to abtain tissue specific expression or stress related histon H3 genes.In this paper,we used the EST Blast method to clone Triticum aestivum histon H3 gene,analyzed its expression changes under salt stress using semi-quatitative RT-PCR,and demonstrated its intron pattern,expression profile and in silico mapping through bioimformatic analysis.1 Cloning and analysis of Triticum aestivum histon H3 geneSeven Triticum aestivum histon H3 gene was cloned by Blasting the Triticum aestivum EST database with the Arabidopsis thaliana homologous protein histom H3 sequence as a querying probe.To examine the accuracy of in silico cloning cDNA sequences,these sequences were cloned by RT-PCR and sequenced.The result confirmed that the in silico cloning cDNA sequences were accurate,and they can derided into two kind:type H3.1 and type H3.22 in silico analysis of Triticum aestivum histon H3 geneIn order to deternine the intron pattern of wheat histon H3 gene,specific primers which were designed for cDNA cloning were also used to amplify genomic DNA, three out of seven genomic sequences of histone H3 genes were abtained,and only one of them had one 198 bp intron,which is a little longer than those homolog genes in other species.In silico expression patterns in different tissues of those seven wheat histon H3 genes were investigated by Unigene database,the results shows that they have no tissue specific expression.Also Graingene database was used to determine their In silico mapping,which shows they were distributed over chromesome 7BL, 4AL and 7AS.3 expression of Triticum aestivum histon H3 gene in response to salt treatmentSemi-quantitative RT-PCR assay was performed to detect the expression pattern of the obtained Triticum aestivum histon H3 gene under salt stress,the result showed that the transcription of one histon H3 gene is accumulated after treated by salt.