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吡咯伯克霍尔德氏菌JK-SH007多聚半乳糖醛酸酶基因的克隆及表达分析(PDF)

《南京林业大学学报(自然科学版)》[ISSN:1000-2006/CN:32-1161/S]

Issue:
2018年04期
Page:
127-133
Column:
研究论文
publishdate:
2018-07-12

Article Info:/Info

Title:
Cloning and expression analysis of polygalacturonase gene(BpPG) from Burkholderia pyrrocinia JK-SH007
Article ID:
1000-2006(2018)04-0127-07
Author(s):
CHEN FeifeiHUANG JinsiWANG XiyunLIU WanhuiYE Jianren*
Co-Innovation Center for the Sustainable Forestry in Southern China, College of Forestry, Nanjing Forestry University, Nanjing 210037, China
Keywords:
Keywords:Burkholderia pyrrocinia BpPG gene molecular cloning expression analysis
Classification number :
Q939.96; S763.1
DOI:
10.3969/j.issn.1000-2006.201708013
Document Code:
A
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.

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Last Update: 2018-07-27