Roh In-Soon, Kim Yong-Chan, Kim Hyo-Jin, Won Sae-Young, Jeong Min-Ju, Kang Hae-Eun, Sohn Hyun-Joo, Jeong Byung-Hoon
Reference Laboratory for CWD, Foreign Animal Disease Division, Animal and Plant Quarantine Agency, Gimcheon, 39660, Republic of Korea.
Korea Zoonosis Research Institute, Jeonbuk National University, 820-120, Hana-ro, Iksan, Jeonbuk, 54531, Republic of Korea.
Mol Biol Rep. 2020 Aug;47(8):6155-6164. doi: 10.1007/s11033-020-05697-9. Epub 2020 Jul 31.
Chronic wasting disease (CWD) is caused by abnormal deleterious prion protein (PrP), and transmissible spongiform encephalopathy occurs in the Cervidae family. In recent studies, the susceptibility of prion disease has been affected by polymorphisms of the prion gene family. However, the study of the prion-related protein gene (PRNT) is rare, and the DNA sequence of this gene was not fully reported in all Cervidae families. In the present study, we amplified and first identified PRNT DNA sequences in the Cervidae family, including red deer, elk, sika deer and Korean water deer, using polymerase chain reaction (PCR). We aligned nucleotide sequences of the PRNT gene and the amino acid sequences of prion-related protein (Prt) protein among several species. In addition, we performed phylogenetic analysis to measure the evolutionary relationships of the PRNT gene in the Cervidae family. Furthermore, we performed homology modeling of the Prt protein using SWISS-MODEL and compared the structure of Prt protein between sheep and the Cervidae family using the Swiss-PdbViewer program. We obtained much longer PRNT sequences of red deer compared to the PRNT gene sequence registered in GenBank. Korean water deer denoted more close evolutionary distances with goats and cattle than the Cervidae family. We found 6 Cervidae family-specific amino acids by the alignment of Prt amino acid sequences. There are significantly different distributions of hydrogen bonds and the atomic distance of the N-terminal tail and C-terminal tail between sheep and the Cervidae family. We also detected the mRNA expression of PRNT gene in 3 tissues investigated. To our knowledge, this report is the first genetic study of the PRNT gene in the Cervidae family.
慢性消耗病（CWD）由异常有害的朊病毒蛋白（PrP）引起，是发生在鹿科动物中的传染性海绵状脑病。在最近的研究中，朊病毒疾病的易感性受到朊病毒基因家族多态性的影响。然而，对朊病毒相关蛋白基因（PRNT）的研究很少，并且该基因的DNA序列在所有鹿科动物中并未得到充分报道。在本研究中，我们使用聚合酶链反应（PCR）扩增并首次鉴定了鹿科动物（包括马鹿、麋鹿、梅花鹿和獐）中的PRNT DNA序列。我们比对了几种物种中PRNT基因的核苷酸序列和朊病毒相关蛋白（Prt）的氨基酸序列。此外，我们进行了系统发育分析以衡量鹿科动物中PRNT基因的进化关系。此外，我们使用SWISS-MODEL对Prt蛋白进行了同源建模，并使用Swiss-PdbViewer程序比较了绵羊和鹿科动物之间Prt蛋白的结构。与GenBank中登记的PRNT基因序列相比，我们获得了更长的马鹿PRNT序列。獐与山羊和牛的进化距离比鹿科动物更近。通过比对Prt氨基酸序列，我们发现了6个鹿科动物特有的氨基酸。绵羊和鹿科动物之间的氢键分布以及N末端尾巴和C末端尾巴的原子距离存在显著差异。我们还检测了所研究的3种组织中PRNT基因的mRNA表达。据我们所知，本报告是鹿科动物中PRNT基因的首次遗传学研究。
