宏基因组学及其研究进展

宏基因组学是近年来发展起来的一门新兴学科,主要技术包括从环境样品中提取微生物混合基因组DNA、利用可培养的宿主菌建立宏基因组文库及筛选目的基因。该技术可以克服传统培养技术的不足,是研究未培养微生物、寻找新功能基因和开发获得新资源的重要新途径。目前宏基因组学已广泛应用于各个领域,并在医药、农业、能源开发、环境修复、生物技术、生物防御等方面有了较深入的研究。笔者对宏基因组学的主要技术方法及其应用研究进展进行了简要综述。     

DOPPLER SONOGRAPHIC EVALUATION OF THE DIGITAL BLOOD FLOW IN HORSES WITH LAMINITIS OR SEPTIC PODODERMATITIS

The aim of this study was to evaluate the use of Doppler ultrasonography to monitor vascular blood flow dynamics in defined diseases of the equine digit in a noninvasive way. Doppler sonography was used to evaluate medial digital artery blood flow in eight horses with septic pododermatitis and four horses with laminitis in comparison with 10 horses of a control group. Doppler sonographic measurement and lameness examinations were performed in lame horses before treatment (day 0) and at 3, 6, and 9 days following treatment. Before treatment, blood flow velocities, arterial diameter, and flow volume were significantly higher in the lame horses, while pulsatility indices (PIs) were significantly lower (P < 0.05). Compared with the control group, there was no significant difference of resistive indices (RI) in the septic pododermatitis group, but these indices were significantly lower in the laminitis group. No significant difference between the two lame groups was recorded. After treatment, blood flow velocities and flow volume decreased significantly and PIs increased significantly in both lame groups, while RI and arterial diameter did not change. No correlation between Doppler parameters and the degree of lameness was found. Doppler sonography can be used to monitor vascular blood flow dynamics in horses suffering from septic pododermatitis and laminitis, while blood flow velocities and PI can serve as parameters for monitoring the course of the disease.     

校企合作共建高职畜牧专业教学资源库的研究与实践

专业教学资源库建设是促进教学改革、提高教学质量的重要切入点,是借助网络信息技术平台,实现优质教学资源共享,为教师教学、学生和社会人员自主学习提供系统资源的综合工程.针对校企合作教学资源库建设研究与实践,探讨了高职畜牧专业教学资源库建设的背景与定位、建设目标与思路、主要建设内容以及问题与思考.     

Utilization of digital differential display to identify differentially expressed genes related to rumen development

This study aimed to identify the genes associated with the development of the rumen epithelium by screening for candidate genes by digital differential display (DDD) in silico. Using DDD in NCBI's UniGene database, expressed sequence tag (EST)‐based gene expression profiles were analyzed in rumen, reticulum, omasum, abomasum and other tissues in cattle. One hundred and ten candidate genes with high expression in the rumen were derived from a library of all tissues. The expression levels of 11 genes in all candidate genes were analyzed in the rumen, reticulum, omasum and abomasum of nine Japanese Black male calves (5‐week‐old pre‐weaning: n = 3; 15‐week‐old weaned calves: n = 6). Among the 11 genes, only 3‐hydroxy‐3‐methylglutaryl‐CoA synthase 2 (HMGCS2), aldo‐keto reductase family 1, member C1‐like (AKR1C1), and fatty acid binding protein 3 (FABP3) showed significant changes in the levels of gene expression in the rumen between the pre‐ and post‐weaning of calves. These results indicate that DDD analysis in silico can be useful for screening candidate genes related to rumen development, and that the changes in expression levels of three genes in the rumen may have been caused by weaning, aging or both. © 2015 Japanese Society of Animal Science     

猪蛔虫感染期幼虫抑制消减cDNA文库的构建

为筛选与线虫感染性相关的基因,本研究以猪蛔虫为对象,构建猪蛔虫感染期幼虫差异表达消减cDNA文库,为研究线虫期特异性发育的分子机制奠定基础。分别提取感染期幼虫和其它各期幼虫及成虫的总RNA,纯化mRNA后,采用Clontech公司PCR-selectTM试剂盒进行反转录合成cDNA并进行抑制消减杂交(SSH),构建猪蛔虫感染期幼虫差异表达的消减cDNA文库,并采用Southern斑点杂交进行消减效率的检测。随机从文库中抽取45个克隆进行测序及在线BLAST分析。试验结果表明,感染期幼虫差异表达的消减cDNA文库具有较强的特异性;在得到的41个表达序列标签(ESTs)中,有40个ESTs与已报道的基因有较高的相似性,主要代表猪蛔虫第三期幼虫基因和成虫头部基因,有1个cDNA片段可能代表新基因。猪蛔虫感染期幼虫差异表达消减cDNA文库的成功构建,为进一步研究幼虫发育差异表达基因的功能奠定了基础。     

猪瘟病毒E2基因噬菌体展示多肽库的构建及表位鉴定

本研究通过建立CSFV噬菌体展示多肽库,并对其进行生物淘选,以期获得E2蛋白上新的抗原表位.选择CSFV石门株(SM)和疫苗株(HCLV)为代表株,采用噬菌体展示技术,以T7select415-1b为载体,分别构建了CSFV SM株和HCLV株E2基因噬菌体展示多肽库(SM-E2库和HCLV-E2库).通过生物淘选和噬菌体原位杂交技术,采用7株猪瘟单克隆抗体和1株猪瘟高免血清分别对构建的SM-E2库和HCLV-E2库进行抗原表位筛选.结果共筛选到了5条与E2蛋白高度同源的序列,在E2蛋白上的同源区域分别为TAVSPTTLR、YYEP、TTWKEYSH、GGQ(V)VK和PDGLPHY.结果表明,TAVSPTTLR、YYEP和TTWKEYSH序列与目前已知E2蛋白表位一致,说明它们是E2蛋白上的优势表位;GGQ(V)VK和PDGLPHY序列与预测表位一致,推测是E2蛋白上潜在的抗原表位.     

表达序列标签（ESTs）及其应用

表达序列标签(ESTs)是指从(ESTcDNA文库中随机挑取克隆并对其3’或5’端进行单轮自动测序所获得的短cDNA库列，一般长度为300～500bp。要有效获取ESTs，必须对cDNA文库进行预处理，处理方法有衰减杂交法、均一化法和差异显示法。在dbEST中收录了大量各种生物的ESTs。ESTs广泛应用于鉴定基因、发现新基因、电子克隆、构建遗传学图谱、制备DNA芯片、分子标记、研究基因的差异表达及检验病原微生物等方面。     

Superficial digital flexor tendonitis in Thoroughbred race horses: outcome following non-surgical treatment and superior check desmotomy

Objective This study documents the results of non-surgical treatment and treatment by superior check desmotomy in Thoroughbred racehorses with superficial digital flexor (SDF) tendonitis. Design A prospective study was made of 124 thoroughbred racehorses with unilateral or bilateral SDF tendonitis. Procedure The flexor tendons were assessed by physical and ultrasonographic examination before treatment, and the lesions detected in affected tendons were characterised according to lesion type, length and cross-sectional area. Ninety three horses were managed non-surgically and 31 by superior check desmotomy. Recurrent or new injuries were defined as injuries affecting a previously injured superficial digital flexor tendon, the contralateral SDF tendon, or the suspensory ligament (interosseous muscle) in either forelimb. Results No statistically significant difference was found in ultrasonographic lesion severity between treatment groups. Horses managed by superior check desmotomy were 1.3 times more likely to complete five or more races than horses managed non-surgically (95% confidence limits 0.93–1.82). Horses treated surgically were 1.2 times more likely to develop recurrent or new injuries after returning to training than horses managed non-surgically (95% CL 0.95–1.55). Horses under-going superior check desmotomy were 5.5 times more likely to develop suspensory desmitis than horses treated non-surgically (95% CL 1.13–26.4). There was no difference in the time to recurrent or new injury between treatment groups. Conclusion There was no statistically significant difference between treatment groups in the proportions of horses able to complete five or more races after an episode of superficial digital flexor tendonitis. Superior check desmotomy did not appear to offer an advantage over non-surgical treatment in preventing recurrent or new injuries in Thoroughbred racehorses. Horses undergoing superior check desmotomy appeared to be at greater risk of developing suspensery ligament injuries than horses managed non-surgically.