Qazi Izhar Hyder, Yuan Ting, Yang Sijia, Angel Christiana, Liu Jiping
Guangdong Provincial Key Lab of Agro-Animal Genomics and Molecular Breeding, College of Animal Science, South China Agricultural University, Guangzhou, China.
Shaheed Benazir Bhutto University of Veterinary and Animal Sciences, Sakrand, Pakistan.
Front Vet Sci. 2024 Jun 28;11:1429169. doi: 10.3389/fvets.2024.1429169. eCollection 2024.
Pebrine, caused by microsporidium , is a devastating disease that causes serious economic damages to the sericulture industry. Studies on development of therapeutic and diagnostic options for managing pebrine in silkworms are very limited. () of microsporidia is an essential gene for their survival and has been exploited as the cellular target of drugs such as fumagillin and its analogues in several microsporidia spp., including Nosema of honeybees.
In the present study, using molecular and bioinformatics tools, we performed in-depth characterization and phylogenetic analyses of MetAP2 of isolated from Guangdong province of China.
The full length of gene sequence of (Guangdong isolate) was found to be 1278 base pairs (bp), including an open reading frame of 1,077 bp, encoding a total of 358 amino acids. The bioinformatics analyses predicted the presence of typical alpha-helix structural elements, and absence of transmembrane domains and signal peptides. Additionally, other characteristics of a stable protein were also predicted. The homology-based 3D models of MetAP2 of (Guangdong isolate) with high accuracy and reliability were developed. The MetAP2 protein was expressed and purified. The observed molecular weight of MetAP2 protein was found to be ~43-45 kDa. The phylogenetic analyses showed that MetAP2 gene and amino acids sequences of (Guangdong isolate) shared a close evolutionary relationship with spp. of wild silkworms, but it was divergent from microsporidian spp. of other insects, spp., , and higher animals including humans. These analyses indicated that the conservation and evolutionary relationships of MetAP2 are closely linked to the species relationships.
This study provides solid foundational information that could be helpful in optimization and development of diagnostic and treatment options for managing the threat of infection in sericulture industry of China.
由微孢子虫引起的微粒子病是一种毁灭性疾病，给养蚕业造成严重经济损失。关于开发治疗和诊断方法来防治家蚕微粒子病的研究非常有限。微孢子虫的甲硫氨酸氨肽酶2（MetAP2）是其生存所必需的基因，在包括蜜蜂微孢子虫在内的几种微孢子虫中，已被用作烟曲霉毒素及其类似物等药物的细胞靶点。
在本研究中，我们使用分子和生物信息学工具，对从中国广东省分离的微孢子虫的MetAP2进行了深入表征和系统发育分析。
发现微孢子虫（广东分离株）的基因序列全长为1278个碱基对（bp），包括一个1077 bp的开放阅读框，共编码358个氨基酸。生物信息学分析预测其存在典型的α-螺旋结构元件，不存在跨膜结构域和信号肽。此外，还预测了该蛋白的其他稳定特性。构建了具有高精度和可靠性的微孢子虫（广东分离株）MetAP2的基于同源性的三维模型。表达并纯化了MetAP2蛋白。观察到MetAP2蛋白的分子量约为43 - 45 kDa。系统发育分析表明，微孢子虫（广东分离株）的MetAP2基因和氨基酸序列与野生家蚕的微孢子虫物种具有密切的进化关系，但与其他昆虫的微孢子虫物种、人类等高等动物的微孢子虫物种不同。这些分析表明，MetAP2的保守性和进化关系与物种关系密切相关。
本研究提供了坚实的基础信息，有助于优化和开发诊断及治疗方法，以应对中国养蚕业中微粒子虫感染的威胁。
