Zhang Qiang, Shao Qian, Guo Yaqiong, Li Na, Li Yu, Su Jiayuan, Xu Rui, Zhang Ziding, Xiao Lihua, Feng Yaoyu
State Key Laboratory of Bioreactor Engineering, School of Resource and Environmental, East China University of Science and Technology, Shanghai, China.
Center for Emerging and Zoonotic Diseases, College of Veterinary Medicine, South China Agricultural University, Guangzhou, China.
Front Microbiol. 2021 Jan 12;11:622203. doi: 10.3389/fmicb.2020.622203. eCollection 2020.
In spp., calcium-dependent protein kinases (CDPKs) are considered promising targets for the development of pharmaceutical interventions. Whole-genome sequencing has revealed the presence of 11 CDPKs in (CpCDPKs). In this study, we expressed recombinant CpCDPK4, CpCDPK5, and CpCDPK6 in . The biological characteristics and functions of these CpCDPKs were examined by using quantitative reverse transcription PCR (qRT-PCR), immunofluorescence microscopy, and an neutralization assay. The expression of the CpCDPK4 gene peaked at 12 h post-infection, the CpCDPK5 gene peaked at 12 and 48 h, and the CpCDPK6 gene peaked at 2-6 h. CpCDPK4 protein was located in the anterior and mid-anterior regions of sporozoites, and CpCDPK5 protein was located over the entire sporozoites, while CpCDPK6 protein was expressed in a spotty pattern. Immune sera of CpCDPK4 and CpCDPK6 exhibited significant inhibitory effects on host cell invasion, while the immune sera of CpCDPK5 had no effects. These differences in protein localization, gene expressions, and neutralizing capacities indicated that the CpCDPK proteins may have different roles during the lifecycle of spp.
在[物种名称]中，钙依赖性蛋白激酶（CDPKs）被认为是药物干预开发的有前景的靶点。全基因组测序揭示了[物种名称]中存在11种CDPKs（CpCDPKs）。在本研究中，我们在[具体表达体系]中表达了重组CpCDPK4、CpCDPK5和CpCDPK6。通过定量逆转录PCR（qRT-PCR）、免疫荧光显微镜和[具体中和试验名称]中和试验检测了这些CpCDPKs的生物学特性和功能。CpCDPK4基因的表达在感染后12小时达到峰值，CpCDPK5基因在12小时和48小时达到峰值，而CpCDPK6基因在2至6小时达到峰值。CpCDPK4蛋白位于子孢子的前部和前中部区域，CpCDPK5蛋白位于整个子孢子上，而CpCDPK6蛋白以斑点状模式表达。CpCDPK4和CpCDPK6的免疫血清对宿主细胞入侵表现出显著的抑制作用，而CpCDPK5的免疫血清则没有作用。这些蛋白质定位、基因表达和中和能力的差异表明，CpCDPK蛋白在[物种名称]的生命周期中可能具有不同的作用。
