Cloning and expression analysis of polygalacturonase gene(BpPG)
from Burkholderia pyrrocinia JK-SH007

doi:10.3969/j.issn.1000-2006.201708013

JOURNAL OF NANJING FORESTRY UNIVERSITY ›› 2018, Vol. 42 ›› Issue (04): 127-133.doi: 10.3969/j.issn.1000-2006.201708013

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Cloning and expression analysis of polygalacturonase gene(BpPG) from Burkholderia pyrrocinia JK-SH007

CHEN Feifei,HUANG Jinsi,WANG Xiyun,LIU Wanhui,YE Jianren^*

Co-Innovation Center for the Sustainable Forestry in Southern China, College of Forestry, Nanjing Forestry University, Nanjing 210037, China

Online:2018-07-27 Published:2018-07-27

Abstract

Abstract: Abstract: 【Objective】In this study, we aimed to isolate the polygalacturonase gene(BpPG)from Burkholderia pyrrocinia JK-SH007 and clarify the biological characteristics of the BpPG in colonizing B. pyrrocinia JK-SH007.【Method】The BpPG was isolated from B. pyrrocinia JK-SH007 by PCR using primers PG-F and PG-F. Bioinformatics and homology analysis were conducted using various online software products and the phylogenetic tree was constructed with MEGA 5.0. The RT-qPCR method was used to analyze differences in the specific expression of the BpPG gene during B. pyrrocinia JK-SH007 colonization of poplar. 【Result】The sequence of the BpPG was 2 001 bp in length, and encoded a protein of 666 amino acids with a molecular weight of 69.23 ku and a theoretical isoelectric point of 8.37. The BpPG was homologous to polygalacturonase(WP 034181566.1), with 99% similarity, and contained six protein binding sites. BpPG is a hydrophilic protein belonging to the glycosyl hydrolase family 28, and lacks a signal peptide. The secondary structures of the protein are comprised mainly of alpha helices, beta turns, extended strands, and random coils, which contain six PbH1 motifs. The predicted tertiary structure was consistent with the secondary structure. RT-qPCR analysis revealed that the relative expression of the BpPG differed during various periods of B. pyrrocinia JK-SH007 colonization of poplar. 【Conclusion】The BpPG isolated from B. pyrrocinia JK-SH007 had a conserved sequence. In addition, the results suggest that the BpPG plays an important role during B. pyrrocinia JK-SH007 colonization of poplar.

CLC Number:

CHEN Feifei,HUANG Jinsi,WANG Xiyun,LIU Wanhui,YE Jianren. Cloning and expression analysis of polygalacturonase gene(BpPG) from Burkholderia pyrrocinia JK-SH007[J]. JOURNAL OF NANJING FORESTRY UNIVERSITY, 2018, 42(04): 127-133.

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Cloning and expression analysis of polygalacturonase gene(BpPG) from Burkholderia pyrrocinia JK-SH007

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