학술논문

Identification of genetic determinants of hemolytic activity of Riemerella anatipestifer using random transposon mutagenesis
Document Type
Academic Journal
Source
Veterinary Research. February 12, 2021, Vol. 52 Issue 1
Subject
Transposons -- Genetic aspects
Genomics -- Genetic aspects
Membrane proteins -- Genetic aspects
Recombinant proteins -- Genetic aspects
Language
English
ISSN
0928-4249
Abstract
Riemerella anatipestifer causes epizootic infectious disease in poultry resulting in serious economic losses especially to the duck industry. In our previous study, R. anatipestifer was found to lyse duck erythrocytes in vitro. In the present study, a random Tn4351 mutagenesis library of hemolytic R. anatipestifer strain SX containing 4000 mutants was constructed to investigate the genetic basis of hemolytic activity. Thirty mutants with reduced hemolytic activity and one with increased hemolytic activity were screened and insertions in 24 genes were identified. Of these genes, four were predicted to encode outer membrane proteins, one encoded a cytoplasmic membrane protein, 11 encoded cytoplasmic proteins, and eight encoded proteins with unknown locations. Based on current annotations of the R. anatipestifer genomes, of the 24 genes, 7 (29.17%) were involved in iron utilization. The hemolytic activities of the complemented strains M2 (pRES-Riean_0790) and M18 (pRES-Riean_0653) were restored, indicating that both Riean_0653 and Riean_0790 are involved in the hemolytic activity of strain SX. However, the recombinant proteins rRiean_0317, rRiean_0790, rRiean_0653, rRiean_1027, rRiean_1143, and rRiean_1561 had no hemolytic activity, suggesting that none were hemolysins. Keywords: Riemerella anatipestifer, Random transposon mutagenesis, Hemolytic activity, Hemolysin, Recombinant protein
Author(s): Bingqing Sun[sup.1,2], Yafei Xue[sup.1], Xiaoli Du[sup.1], Xiaohua He[sup.1], Zuocheng Zou[sup.1], Xiangqiang Tian[sup.1], Zhonghao Hu[sup.1], Haoyang Liu[sup.1], Nazrul Islam[sup.1] and Qinghai Hu[sup.1] Introduction Riemerella anatipestifer, the type species of the [...]