Cloning and Detection of Equine Histidine-Rich Glycoprotein

被引：3

作者：

Muko, Ryo ^{[1
]}

Amagai, Yosuke ^{[2
]}

Matsuda, Kenshiro ^{[3
]}

Matsuda, Hiroshi ^{[1
,3
]}

Tanaka, Akane ^{[1
,4
]}

机构：

[1] Tokyo Univ Agr & Technol, Grad Sch Bioapplicat & Syst Engn, Adv Hlth Sci, Fuchu, Tokyo, Japan

[2] Tokyo Metropoltan Inst Med Sci, Japan Soc Promot Sci, Dept Mol Med Res, Setagaya Ku, Tokyo, Japan

[3] Tokyo Univ Agr & Technol, Inst Agr, Div Anim Life Sci, Lab Vet Mol Pathol & Therapeut, Fuchu, Tokyo, Japan

[4] Tokyo Univ Agr & Technol, Inst Agr, Div Anim Life Sci, Lab Comparat Anim Med, Fuchu, Tokyo, Japan

来源：

JOURNAL OF EQUINE VETERINARY SCIENCE | 2019年 / 73卷

关键词：

Horses; Histidine-rich glycoprotein; Cloning; RT-PCR; SIRS; Western blotting; PLASMA-PROTEIN; HIGH-AFFINITY; BINDING; PLASMINOGEN; FIBRINOLYSIS; DEFINITIONS; COAGULATION; SEPSIS; ACID;

D O I：

10.1016/j.jevs.2018.12.009

中图分类号：

S85 [动物医学（兽医学）];

学科分类号：

0906 ;

摘要：

Histidine-rich glycoprotein (HRG) is a plasma glycoprotein that is presumed to be involved in immune response, coagulation, and fibrinolysis. Recent studies have revealed that the blood HRG levels are markedly decreased in the murine model for sepsis, and that recombinant HRG injections significantly improved their survival rate. Although sepsis is a common secondary condition in horses, changes in the blood HRG levels during critical diseases in horses have been unexplored. To elucidate the association of HRG with certain diseases in horses, the establishment of a procedure for the detection of HRG is necessary. In the present study, we investigated HRG messenger RNA (mRNA) expression by real-time polymerase chain reaction using complementary DNA (cDNA) generated from various organ samples of Thoroughbred horses and determined that HRG mRNA was expressed in the liver. Consequently, HRG was cloned using cDNA from the liver samples, and the whole sequence of equine HRG and the amino acid sequence of HRG were identified. Since HRG comprises a significant number of histidine residues, we purified equine HRG in serum using a nickel resin. In addition, the purified equine HRG was detected by Western blot analysis. We also generated recombinant equine HRG, and cross-reactivity of the anti-body was confirmed. We demonstrated the complete sequence of the equine HRG and detected serum HRG in healthy horses for the first time. This information may prove valuable for future investigations of serious inflammatory diseases in horses, such as the systemic inflammatory response syndrome. (C) 2018 Elsevier Inc. All rights reserved.

引用

页码：121 / 126

页数：6

共 23 条

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