Abstract：
Fowl cholera is an infectious disease that affects both poultry and wild birds, characterized by hemorrhagic and septicemic symptoms, caused by Pasteurella multocida (P. multocida), and leading to substantial economic losses in the poultry sector. The development of genetic engineering vaccines against avian P. multocida encountered early-stage challenges due to the limited availability of effective gene editing tools. Presently, NgAgoDM-enhanced homologous recombination stands as a potent technique for achieving efficient gene knockout in avian P. multocida. Hence, this study employed NgAgoDM-enhanced homologous recombination to target and knockout hyaE (239-359aa), hyaD, hexABC, and hexD, denoted as ΔhyaE (239-359aa), ΔhyaD, ΔhexABC, and ΔhexD, respectively. Additionally, we generated a hyaD recovery strain with two point mutations, designated as mhyaD. Thus, this study systematically examined the impact of capsular synthetic gene clusters on the pathogenicity of P. multocida. Moreover, the study demonstrated the critical role of hyaD activity in the virulence of avian P. multocida. This study offers novel insights for enhancing attenuated vaccines further.
摘要：
家禽霍乱是一种传染病，影响家禽和野生鸟类，以出血和败血症症状为特征，由多杀巴斯德氏菌(P.multocida),并导致家禽行业的重大经济损失。由于有效基因编辑工具的可用性有限，针对禽多杀性疟原虫的基因工程疫苗的开发遇到了早期挑战。目前,NgAgoDM增强的同源重组是一种有效的技术，可以在禽类多杀性疟原虫中实现有效的基因敲除。因此，本研究采用NgAgoDM增强同源重组靶向和敲除hyaE（239-359aa），hyaD,hexABC,和十六进制,表示为ΔhyaE(239-359aa)，ΔhyaD,ΔhexABC,和ΔhexD，分别。此外,我们产生了具有两个点突变的hyaD恢复菌株，指定为mhyaD。因此，本研究系统研究了荚膜合成基因簇对多杀性疟原虫致病性的影响。此外，该研究证明了hyaD活性在禽类多杀性疟原虫毒力中的关键作用。这项研究为进一步增强减毒疫苗提供了新的见解。
