Construction and Identification of Yeast Surface Display Libraries of Ovis aries DRA Gene Exon 2     

XU Wan-yun  WANG Hui-min  WANG Wen-lun  HU Meng-wei  YAN Guo  LI Jian-hua  GAO Jian-feng 《中国畜牧兽医》2016,43(3):568-576

The specific primers were designed according to Ovis aries DRA gene sequence deposited in GenBank and the multiple cloning site of the plasmid pYD1,which was a vector used for protein surface display on Saccharomyces cerevisiae.The gene encoding DRA was amplified by PCR using the genomic RNA of Ovis aries.The 762 bp fragment was cloned and released in GenBank and registration number was KR422362.The PCR product was inserted into the yeast surface display plasmid vector pYD1 by double enzyme digestion.It was indicated that DRA gene was successfully integrated into the genome.Dot mutation was made at both ends of exon 2 in DRA gene for making restriction enzyme cutting site and design the exon 2 specific primers according to mutated Ovis aries DRA gene sequence.Sequenced exon 2 amplification products based on DNA pooling of sheep large sample template was analyzed the polymorphic loci.The polymorphic exon 2 246 bp fragment was obtained by double enzyme digestion and connected to surface display restructuring mutation carriers pYD1-DRA by the same double enzyme digestion,and then we successfully constructed yeast surface display libraries.We transformed it into Saccharomyces cerevisiae EBY100 cell.Yeast monoclone was identified by PCR amplification and sequencing,and we confirmed that DRA gene had been integrated into Saccharomyces cerevisiae genome.After galactose induced,it was detected that DRA gene library had been successfully demonstrated on the yeast cell surface under the fluorescence microscope by immunofluorescence method.  相似文献   

内质网应激偶联炎性反应研究进展     

廖轶  崔永勇  刘春法  杨利峰  赵德明  周向梅 《动物医学进展》2016,(7):89-93

内质网是真核细胞蛋白合成和折叠的主要细胞器,当内质网内蛋白合成或折叠负担增加,引起未折叠或错误折叠蛋白增多时,可激活内质网几条特定信号通路,启动未折叠蛋白反应,这对维持细胞稳态有重要意义。越来越多研究表明,多种炎性反应疾病与内质网应激有密切联系。一方面,内质网应激引起的未折叠蛋白反应可以诱发或者抑制炎症,另一方面,炎性反应也能影响蛋白折叠,从而促进或缓解内质网应激。2型糖尿病、肿瘤和动脉粥样硬化等多种重大疾病的病理机制都涉及内质网应激与炎性反应的相互作用,对该问题的深入研究不仅能加深人们对这些疾病发病机制的理解,也有助于相关药物的研发。  相似文献   

响应面法优化杜仲叶提取物的制备工艺     

侯伟峰  郭超  范云鹏  麻武仁  宋晓平 《动物医学进展》2016,(11):74-77

优化杜仲叶提取物制备的工艺条件,在单因素试验基础上,以杜仲叶提取物浸膏得率和绿原酸转移率为评价指标,采用Box-Behnken中心组合设计法优化杜仲叶提取物制备的工艺条件。杜仲叶提取物制备的最佳工艺条件为乙醇浓度500mL/L,液料比10∶1(mL/g),提取温度97℃,提取4次,每次30min;在此最佳工艺条件下,杜仲叶提取物浸膏得率和绿原酸转移率综合评分均值为99.08,与预测值基本一致。响应面法(RSM)优化的杜仲叶提取物制备工艺稳定可靠。  相似文献   

Increased humoral antibody response of foot-and-mouth disease virus vaccine in growing pigs pre-treated with poly-γ-glutamic acid     

Jee-Hoon Lee  Ik-Jae Kang  A-Reum Kim  You-Sun Noh  Hee-Chun Chung  Bong-Kyun Park 《Journal of veterinary science (Suw?n-si, Korea)》2016,17(2):253-256

This study was conducted to determine if humoral antibody response of foot-and-mouth disease (FMD) vaccine improved in 8-week-old growing pigs born to well-vaccinated sows pre-treated with 60 mg of poly-γ-glutamic acid (γ-PGA) three days before vaccination. Antibody against FMD virus serotype O was measured 0, 2, 4 and 6 weeks post-vaccination, using a PrioCHECK FMDV type O ELISA kit. The results showed that positive antibody reactions against FMDV serotype O antigen among a component of the vaccine significantly increased in response to pre-injection with γ-PGA.  相似文献   

Best practice recommendations for prehospital veterinary care of dogs and cats          下载免费PDF全文

Rita M. Hanel DVM  DACVIM  DACVECC  Lee Palmer DVM  MS  DACVECC  NREMT‐T  WEMT  CCRP  Janice Baker DVM  DACVPM  Jo‐Anne Brenner BA  EMT‐I  EMT‐T  David Dorman DVM  PhD  DABVT  John C. Gicking DVM  DACVECC  Brian Gilger DVM  MS  DACVO  DABT  Cynthia M. Otto DVM  PhD  DACVECC  DACVSMR  Elizabeth Rozanski DVM  DACVECC  DACVIM  Brian Trumpatori DVM 《Journal of Veterinary Emergency and Critical Care》2016,26(2):166-233

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Tear ferning test in horses and its correlation with ocular surface evaluation          下载免费PDF全文

Laura R. Silva  Anderson F. Gouveia  Cleyber J. T. de Fátima  Letícia B. Oliveira  Janildo L. Reis Jr.  Renato F. Ferreira  Concepta M. Pimentel  Paula D. Galera 《Veterinary ophthalmology》2016,19(2):117-123

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Characterization of tuberculous granulomas in different stages of progression and associated tertiary lymphoid tissue in goats experimentally infected with Mycobacterium avium subsp. hominissuis     

《Comparative immunology, microbiology and infectious diseases》2016

Oral infection of goats with Mycobacterium avium subsp. hominissuis (MAH) resulted in a large variety of granulomas in organized gut-associated lymphatic tissues and intestinal lymph nodes. To characterize the cellular composition of granulomas, CD4⁺, CD8⁺, γδ, B lymphocytes and plasma, CD25⁺, CD68⁺, MHC-II⁺, Ki67⁺ and endothelial cells were labeled in consecutive frozen sections by immunohistochemistry and acid fast bacilli (AFB) by Kinyoun stain. Granulomas with extensive necrosis, little mineralization and variable numbers of AFB surrounded by many CD4⁺ T cells, but only few epitheloid macrophages were observed in severely sick goats at 2–3 mpi. They were interpreted as exuberant immune reaction. Organized granulomas with very few AFB were seen in clinically healthy goats at 13 mpi. The necrotic cores were surrounded by a zone of granulomatous infiltrate with many epitheloid macrophages and few lymphocytes. This zone was initially wide and highly vascularized and became progressively smaller. It was enclosed by an increasing layer of connective tissue. All organized granulomas were surrounded by compartimentalized tertiary lymphoid tissue. The granulomas in experimental infection of goats with MAH reflect the heterogeneity of lesions seen in mycobacterial infections of humans and ruminants and are therefore valuable for comparative research.  相似文献   

Regulatory verification on safe use of cytotoxic drugs in veterinary clinics and animal hospitals          下载免费PDF全文

V Fung  M Seneviratne 《Australian veterinary journal》2016,94(11):400-404

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Lungeing on hard and soft surfaces: Movement symmetry of trotting horses considered sound by their owners          下载免费PDF全文

T. Pfau  C. Jennings  H. Mitchell  E. Olsen  A. Walker  A. Egenvall  S. Tröster  R. Weller  M. Rhodin 《Equine veterinary journal》2016,48(1):83-89

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Neurologic Deficits Including Auditory Loss and Recovery of Function in Horses with Temporohyoid Osteoarthropathy          下载免费PDF全文

M. Aleman  M. Spriet  D.C. Williams  J.E. Nieto 《Journal of veterinary internal medicine / American College of Veterinary Internal Medicine》2016,30(1):282-288

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