Abstract: To clarify the role of chitinase in the process of pear resistance disease, the total RNA of Yali was extracted and specifi c primers were designed to amplify the coding region of PbChi Ⅱ protein by RT-PCR,analysed the expression pattern of PbChi Ⅱ after treatment with pathogens. The prokaryotic expression vector pET30a-PbChi Ⅱ was constructed and the recombinant protein was expressed in E.coli BL21 (DE3). The recombinant protein growth ability was analyzed under abiotic stress. The results showed that the length of PbChi Ⅱ (GenBank accession Number:KP876485) gene was 969 bp,Quantitative Real-time PCR (qRT-PCR) analysis revealed that the expression of PbChi Ⅱ was regulated by pathogens,and enhanced up to the peak at 48 h after treatment with Venturia nashicola during 96 h. The PbChi Ⅱ gene was successfully subcloned into the expression vector pET30a. SDS-PAGE analysis showed that a specific recombinant protein of approximately 35.53 kDa was produced in the BL21 (DE3) with the prokaryotic expression vector pET30a-PbChi Ⅱ in 37℃ with 1.0 mmol/L IPTG for 2 hours. This protein enhanced the stress of isolate with NaCl,CuCl₂,CdCl₂ and ZnSO₄. This research provided basic references for further study the function of PbChi Ⅱ.

Key words: Pyrus bretschneideri Yali, Chitinase gene, Prokaryotic expression, Abiotic stresses, Expression pattern

CLC Number: 

• Q78