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Cloning and Prokaryotic Expression of Chitinase Ⅲ Gene(PbChi Ⅲ)from Pyrus bretschneideri Yali
Pages: 1301-1309
Year: Issue:  10
Journal: Journal of Agricultural Biotechnology

Keyword:  Pyrus bretschneideri YaliChitinase Ⅲ gene(ChiⅢ)Expression patternProkaryotic expression;
Abstract: Chitinase(Chi) which is an important pathogenesis-related protein can degrade cell wall components of fungal pathogens,and be induced by salicylic acid(SA),ethylene,plant pathogenic fungi,bacteria and virus.To explore the function of chitinaseⅢ(ChiⅢ) gene in Pyrus bretschneideri Yali,the CDS of ChiⅢ was cloned from Yali which was named Pb ChiⅢ(Gen Bank accession No.KJ872675),and the CDS length of Pb ChiⅢ was 897 bp.Homology analysis showed that the amino acid sequence of Pb ChiⅢ was highly homologous with Pyrus Huangguan(ACM45715.1),and the nucleotide acid sequence had the closest relationship with Huangguan(FJ589785.1).Quantitative Real-time PCR(q RT-PCR) analysis revealed that Pb ChiⅢ highly expressed in root and fruit,and its expression had extremely significant difference compared with stem and leave(P<0.01).Expression of Pb ChiⅢ was regulated by SA,and enhanced up to the peak at12 h after treatment with SA during 96 h,and had extremely significant difference compared with control(P<0.01).Pb ChiⅢ protein successfully expressed by prokaryotic expression system and a specific protein band of31 k D was successfully induced.The CDS sequence of Pb Chi Ⅲ gene was firstly obtained from Yali and researched the expression of Pb ChiⅢ in different tissues and the expression characteristic after treatment with SA.This study provided informative references for further study of the function of Pb ChiⅢ.
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