鸽源Ⅰ型菌毛簇的克隆与表达及功能蛋白氨基酸序列特征分析。

Cloning and Expression of Pigeon-Derived Type 1 Pilus Clusters and Analysis of Amino Acid Sequence Characteristics of Functional Proteins.

机构信息

School of Animal Science and Food Engineering, Jinling Institute of Technology, Nanjing 210046, China.

College of Veterinary Medicine, Yangzhou University, Yangzhou 225009, China.

出版信息

Genes (Basel). 2024 Sep 26;15(10):1253. doi: 10.3390/genes15101253.

DOI:10.3390/genes15101253

PMID:39457377

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC11508147/

Abstract

BACKGROUND

Type 1 pili, as an important virulence factor of , has certain homology between APEC and UPEC, but the homology degree is not clear enough.

OBJECTIVES

This study aims to compare the homology between them.

METHODS

The recombinant bacteria were constructed by homologous recombination. The pili were observed by TEM, and the hemagglutination characteristics were determined by MHSA. The complete gene sequence was determined by sequencing, and the amino acid sequences of the functional proteins of type 1 pili of APEC and UPEC were compared.

RESULTS

TEM showed that they could express pili, which were slender, straight, and dense. Stable-pUC- has MHSA but stable-pUC- does not. The amino acid sequence similarity of FimB of NJ05 and UPEC was 98.8%, FimE was 99.4%, and the similarity between them was 51.5%. Compared with UPEC's type 1 pili FimC and FimD sequences, the similarity was 99.52% and 87.8%, respectively. The amino acid sequence of FimA of NJ05 was 89-96%, similar to UPEC, and the N-terminal and C-terminal amino acid sequences were exactly the same. The gene sequence and amino acid sequence similarity of FimH between them were both above 99%. The similarity of the pilus binding domain of FimH was 52.8%, but only 27.6% in the receptor binding domain. A few of the same amino acid residues were found in the corresponding regions of FimA, FimF, FimG, and FimH.

CONCLUSIONS

The type 1 pili of APEC and UPEC come from the same origin, which is helpful to further reveal the pathogenic mechanism of infection in the poultry respiratory tract.

摘要

背景

Ⅰ型菌毛作为的重要毒力因子，在 APEC 和 UPEC 之间具有一定的同源性，但同源程度尚不清楚。

目的

本研究旨在比较它们之间的同源性。

方法

通过同源重组构建重组菌，TEM 观察菌毛，MHSA 测定其血凝特性，测序确定完整基因序列，比较 APEC 和 UPEC Ⅰ型菌毛功能蛋白的氨基酸序列。

结果

TEM 显示它们可以表达菌毛，菌毛细长、直、密集。稳定-pUC-具有 MHSA，但稳定-pUC-没有。NJ05 和 UPEC 的 FimB 氨基酸序列相似性为 98.8%，FimE 为 99.4%，相似性为 51.5%。与 UPEC 的Ⅰ型菌毛 FimC 和 FimD 序列相比，相似性分别为 99.52%和 87.8%。NJ05 的 FimA 氨基酸序列为 89-96%，与 UPEC 相似，N 端和 C 端氨基酸序列完全相同。它们之间 FimH 的基因序列和氨基酸序列相似性均在 99%以上。FimH 菌毛结合域的相似性为 52.8%，而受体结合域仅为 27.6%。在 FimA、FimF、FimG 和 FimH 的相应区域发现了几个相同的氨基酸残基。