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1.
采用RT-PCR扩增和基因克隆技术鉴定断奶仔猪血管紧张素转化酶2(ACE2)基因序列,并与公布的猪以及其他物种的ACE2基因进行同源性比对.结果,成功克隆出了仔猪ACE2部分序列,该核苷酸序列与GenBank上公布的野猪ACE2核苷酸序列同源性达到98％,与欧洲牛的同源性为87.8％,与人的同源性为78.8％,与禽类(鸡)的同源性较低.遗传进化分析发现该扩增片段与牛的遗传距离最近,与鸡则处在完全不同的2个分枝上.氨基酸进化与基因遗传进化结果一致.本研究为仔猪ACE2的有效表达、生物学活性及其在苏姜猪中作用的研究奠定基础. 相似文献
2.
试验探讨了北京鸭β-catenin基因的生物学功能,为进一步研究该基因对鸭皮肤毛囊的影响奠定基础。以北京鸭胚胎皮肤组织为材料,根据GenBank中发表的鸡、鹅等物种的β-catenin基因序列,设计并合成4对引物,采用RT-PCR方法,克隆北京鸭β-catenincDNA,并运用DNAMAN软件及在线工具对所得到的序列进行生物信息学分析。结果表明:北京鸭β-catenin基因cDNA长度为2996bp(Genbank登录号为FJ169885),包含由2346个碱基组成的编码区(CDs),有起始密码子ATG和终止密码子TAA,编码781个氨基酸;北京鸭β-catenin基因核苷酸序列与鸡、鹅、中华鳖、人和家猪相应区段(CDs)的同源性分别为95.06%、94.12%、90.11%、84.83%、84.31%;其氨基酸序列与鸡、鹅、中华鳖、猪、人的氨基酸同源性达99%以上,表明在进化关系上,β-catenin氨基酸序列有很高的保守性。 相似文献
3.
《黑龙江畜牧兽医》2017,(15)
为了进行猪A-FABP基因的克隆及不同器官组织中的差异表达,试验采用RT-PCR方法从长白猪背最长肌中克隆A-FABP基因的cDNA序列并进行测序比对,分别与人、牛、山羊、家鼠及鸡进行同源性比较,应用半定量RT-PCR方法检测猪心脏、肝脏、脾脏、肺脏、肾脏、背最长肌、腿肌中A-FABP基因的表达。结果表明:成功克隆得到A-FABP基因的cDNA序列,且比对结果为100%,该序列与人、牛、羊的同源性分别为90.98%、89.97%、89.97%,与鸡的同源性为73.68%。半定量RT-PCR结果为A-FABP基因在心脏、肝脏、脾脏、肺脏、肾脏、背最长肌、腿肌器官组织中均有表达,其中背最长肌和腿肌中A-FABP基因的表达量最高,肺脏内的表达量最低,A-FABP基因在动物进化中具有高度保守性,该基因在猪不同器官组织中的表达具有差异性。 相似文献
4.
《畜牧与兽医》2020,(3):38-44
血管紧张素转换酶2(Angiotensin converting enzyme 2,ACE2)作为血管紧张素系统(Renin-Angiotensin System,RAS)的重要一员,对鸡ACE2的研究尚没有开展。本研究以白羽肉鸡为研究对象,从白羽肉鸡空肠组织中扩增出ACE2基因编码区的特异性片段,TA克隆到pMD-19T载体并转到大肠杆菌DH5α感受态细胞,对生长出的单菌落进行蓝白斑筛选、菌液PCR、酶切验证后测序,对所得序列进行了生物信息学分析。结果:本研究明确了ACE2在白羽肉鸡体内有存在,分布表达有组织特异性,在消化系统及肾脏有较高表达。并且首次成功克隆了白羽肉鸡ACE2全基因序列,共包括2,444个核苷酸,编码808个氨基酸残基,同源性及遗传进化结果一致。该基因编码ACE2蛋白属不稳定、亲水性蛋白,蛋白信号肽序列位于第1-17aa之间,蛋白跨膜螺旋预测为I型跨膜蛋白。研究所得序列已上传GenBank(基因序列号为MK560199),为ACE2在肉鸡及禽类方面的研究提供了基础资料。 相似文献
5.
利用设计的1对特异性引物,通过RT-PCR方法扩增出4株鸡传染性支气管炎病毒(IBV)安徽地方分离株膜蛋白M基因全长片段并进行了克隆测序。将各IBV安徽地方分离株与GenBank中注册的一些毒株M基因核苷酸序列及推导的氨基酸序列进行比较和系统进化关系分析,发现毒株间核苷酸序列同源性为88.5%~100%,其相应的氨基酸序列同源性为90.3%~100%;不同毒株间存着重组、缺失、插入及点突变等变异,从ATG至第140 bp区段的核苷酸序列变异频率最高;4株分离毒株属于同一个进化群的2个不同进化亚群,与我国常用疫苗毒株H120、M41和W 93不属同一个进化亚群。 相似文献
6.
通过PCR反应扩增7株鸡奇异变形杆菌和1株兔奇异变形杆菌的23SrRNA基因片段(1045bp),经克隆测序,用DNA Star分析软件将所获得的序列与GenBank中收录的奇异变形杆菌、普通变形杆菌以及其他相近属的23S rRNA基因序列进行同源性比较,并由此构建奇异变形杆菌系统进化发生树。结果表示,本实验室保存的7株鸡奇异变形杆菌核苷酸序列与GenBank中收录的奇异变形杆菌核苷酸序列同源性为99.4%～99.8%,与兔奇异变形杆菌核苷酸序列同源性为98.8%～99.3%,与普通变形杆菌的核苷酸序列同源性为95.4%～96.2%,而与其他相近属同源性只有92.9%～93.4%。结果表明,23S rRNA基因序列分析可以作为鉴定奇异变形杆菌的一种快速、简便的方法。 相似文献
7.
马的ACE基因多态性的探索与研究 总被引：1，自引：0，他引：1
提取马血的总DNA,参照Genbank中人、羊、兔的ACE基因中的16,17外显子序列,设计1对保守引物p1,应用PCR技术扩增出目的基因,将其克隆到PMD19-T载体中进行测序,并用DNAStar软件进行序列分析。结果显示,扩增出的目的基因长度为345bp,与人的ACE基因16～17外显子(11576～12060bp)序列同源性达到82.3%,证明扩增出的基因包括了马的ACE基因16内含子序列。再参照此序列,设计特异性引物p2,对5种马的ACE基因进行PCR扩增,与人的ACE基因多态性的位点进行分析比较。结果证明,马在此位点上不存在多态性。该试验为马ACE耐力基因多态性的继续研究提供了基础资料,为赛马的选材提供了理论依据。 相似文献
8.
《畜牧与兽医》2016,(3):32-36
从福建某鸡场发病鸡的气管和肾脏组织中分离到1株病毒,通过病毒对SPF鸡胚的致病性特征、血凝特性、电镜观察及RT-PCR鉴定等方面的研究,证实该病毒为鸡传染性支气管炎病毒(IBV),并命名为GZ毒株。通过RT-PCR对该毒株的3CLpro基因片段进行扩增,随后进行克隆与测序。该序列与参考毒株的核苷酸序列分析显示,GZ株的3CLpro基因片段与参考毒株的同源性为85.9%~98.5%,与Beaudette毒株核苷酸序列同源性最高(98.5%),而与H120株同源性为89.7%,与LX4株同源性最低(85.9%)。遗传演化分析显示,GZ分离株与参考株可以划分为3个群,GZ分离株与Beaudette毒株处于同一分支,而与LX4、BJ及Massachusetts毒株亲缘关系较远。 相似文献
9.
鸡α干扰素基因的克隆和鉴定 总被引：14，自引：0，他引：14
采用反转录-聚合酶链反应技术，从传染性囊病病毒感染的鸡胚成纤维细胞中扩增出石岐杂鸡的α干扰素（spzChIFN-α)基因，将包括sqzChIFN-α编码区的cDNA片断克隆到pBssK载体中，并对该片断进行序列测定，核苷酸序列测定结果表明克隆的ChIFN-α基因cDNA片段长度为724bp，和其他ChIFN-α基因的同源性在9左.1%以上，与其它α食干扰素基因的同源性在67.4%以上，其中与鸭α干扰素基因的同源性为67.4%，与火鸡α干扰素基因的同源性为89.8%，推导的石岐杂鸡α干扰素氨基酸序列与其它禽类的α干扰素的同源性在49.2%以上，其中与白来航鸡α干扰素的同源性为96.9%-99%，与鸭和火鸡α干扰素同源性分别为49.2%和80.7%。 相似文献
10.
利用PCR方法首次克隆了鸡Cacnals基因5'一部分调控区和exon1共1 863bp的序列,该序列已提交GenBank,登录号为GQ184725.同源性比对发现,鸡Cacnals基因exon1与哺乳动物同源性较高,其核苷酸相似性在72.4%～77.0%,氨基酸相似性在66.0%～2.0%.鸡Cacnals基因exon1较哺乳动物插入了9个碱基,分别编码1个高疏水性的缬氨酸和2个高亲水性的天冬氨酸和赖氨酸.Cacnals基因部分序列的克隆,可为下一步在鸡上开展此基因与肉质和屠体性状的关联性研究奠定基础. 相似文献
11.
中国麻鸭血管紧张素转化酶2(ACE2)的基因克隆及基因结构的序列解析 总被引：1，自引：1，他引：0
血管紧张素转化酶2（ACE2）被发现以来，其病理生理功能，特别是作为SARS、COVID-19等冠状病毒侵入细胞的功能性受体，显示了巨大的潜能，明确其在不同物种的基因序列及结构可为冠状病毒感染机制研究提供依据。本研究以中国麻鸭为研究对象，利用RT-PCR和Western blot检测ACE2在中国麻鸭各组织中的表达，然后用PCR技术，扩增出中国麻鸭ACE2基因的完整ORF序列，再TA克隆至pMD-19T载体中进行测序，对所得序列进行生物信息学解析。结果证实，ACE2在中国麻鸭心、肝、肺、肾等组织中均有表达。基因克隆结果显示，该中国麻鸭ACE2基因的CDS序列全长为2 435 bp，编码805个氨基酸残基，与人ACE2核苷酸序列和氨基酸序列同源性分别为66.2%和66.4%，且处在不同进化树分支上。通过对与SARS病毒S蛋白结合有关的18个关键氨基酸残基分析发现，中国麻鸭除第330和353位氨基酸外，其余均与人不相同。结构分析发现，中国麻鸭ACE2为Ⅰ型跨膜蛋白，有多处N-糖基化位点。研究首次获得中国麻鸭ACE2基因的完整ORF序列及相关基础资料，所得序列已上传GenBank并被成功收录，研究结果为开展ACE2在鸭上的功能研究提供了理论依据。 相似文献
12.
B. M. Pressler L. E. Williams J. A. Ramos-Vara K. I. Anderson 《Journal of veterinary internal medicine / American College of Veterinary Internal Medicine》2009,23(3):592-597
Background: Similarities in human and canine renal cell carcinoma (RCC) epidemiology and biologic behavior suggest that molecular mechanisms of tumorigenesis may be similar in both species. Approximately 75% of RCC in people are of the clear cell subtype, up to 85% of which are associated with mutation of the von Hippel-Lindau ( VHL ) gene. The canine VHL coding deoxyribonucleic acid (DNA) shares 90% identity with the human VHL gene.
Objective: To determine whether or not RCC in dogs are associated with VHL mutations, and if so determine the prevalence, type, and location of these mutations.
Animals: Thirteen dogs with RCC, 2 dogs with primary renal sarcomas, and 10 dogs without neoplastic kidney disease.
Methods: DNA was extracted from paraffin-embedded RCC tissue; DNA extracts from paraffin-embedded and snap-frozen nonneoplastic canine kidneys and canine whole blood were used as negative controls. Polymerase chain reaction and sequencing of the 3 VHL exons was performed, and results compared with the accessioned canine sequence.
Results: All VHL exons were amplified from 9 of 13 canine RCC samples, both renal sarcomas, 8 of 10 nonneoplastic kidney samples, and canine whole blood; only exon 2 could be amplified from 2 RCC samples. Mutations were not identified in any exons. A maximal prevalence of 33.6% for VHL mutations in canine RCC was determined.
Conclusion and Clinical Importance: Although similarities between canine and human RCC merit further investigation of the dog as a model for some subtypes of renal tumors, the lower prevalence of VHL mutations suggests that oncogenesis in these 2 species differs. 相似文献
Objective: To determine whether or not RCC in dogs are associated with VHL mutations, and if so determine the prevalence, type, and location of these mutations.
Animals: Thirteen dogs with RCC, 2 dogs with primary renal sarcomas, and 10 dogs without neoplastic kidney disease.
Methods: DNA was extracted from paraffin-embedded RCC tissue; DNA extracts from paraffin-embedded and snap-frozen nonneoplastic canine kidneys and canine whole blood were used as negative controls. Polymerase chain reaction and sequencing of the 3 VHL exons was performed, and results compared with the accessioned canine sequence.
Results: All VHL exons were amplified from 9 of 13 canine RCC samples, both renal sarcomas, 8 of 10 nonneoplastic kidney samples, and canine whole blood; only exon 2 could be amplified from 2 RCC samples. Mutations were not identified in any exons. A maximal prevalence of 33.6% for VHL mutations in canine RCC was determined.
Conclusion and Clinical Importance: Although similarities between canine and human RCC merit further investigation of the dog as a model for some subtypes of renal tumors, the lower prevalence of VHL mutations suggests that oncogenesis in these 2 species differs. 相似文献
13.
Kitagawa H Eguchi T Kitoh K Ohba Y Kondo M Nakano M Sasaki Y 《The Journal of veterinary medical science / the Japanese Society of Veterinary Science》2000,62(2):179-185
In order to examine the safety of an angiotensin-converting enzyme (ACE) inhibitor in dogs with impaired renal excretion route, benazepril was administered orally, and plasma concentrations of benazeprilat, the active metabolite of benazepril, were determined in dogs with renal mass reduction (1/4th kidney) created by right-side nephrectomy and ligation of branches of the left renal arteries. Five dogs were administered benazepril orally at a given dose (0.5 mg/kg body weight) and 4 other dogs received 20 times that dose (10 mg/kg body weight) once daily for 15 consecutive days before (intact kidney period) and after (1/4th kidney period) creation of kidney impairment. Six control dogs received surgical treatment, but no drug. After creating a 1/4th kidney, plasma urea nitrogen and creatinine concentrations increased to approximately 30 mg/dl and 2.0 mg/dl, respectively, and renal plasma flow and glomerular filtration rate decreased to 37% and 30% of pre-treatment values, respectively. However, these parameters did not change significantly during the 1/4th kidney period both in the 0.5 mg/kg and 10 mg/kg groups. In the 0.5 mg/kg group, plasma benazeprilat concentrations increased to approximately 20 ng/ml to 340 ng/ml 2 hr after each administration, and there were no significant differences between the plasma benazeprilat concentrations during the intact and 1/4th kidney periods. In the 10 mg/kg group, plasma benazeprilat concentrations varied in the individual dog, but did not increase with the days of administration, and were not significantly different on each administration day between the intact and 1/4th kidney periods in either dose group. The AUCs(0-24) of plasma benazeprilat concentrations determined on the 15th administration day were not different between the intact and 1/4th kidney periods in dogs of either dose group. Plasma ACE activities decreased after drug administration in dogs of both groups. Benazepril seemed to have a high safety, and the adjustment of dosage regimen might not be needed in dogs with mild to moderate renal function impairment because the drug was excreted both from the kidneys and liver. 相似文献
14.
血管紧张素转换酶2(ACE2)是首先克隆出的人类血管紧张素转换酶(ACE)的同源基因,是肾素-血管紧张素系统(RAS)的关键调节因子,其主要通过水解血管紧张素Ⅱ(AngⅡ)生成血管紧张素1-7[Ang(1-7)]从而发挥舒张血管、抗炎、抗增殖等作用。近期研究发现,ACE2不仅对心血管和肾功能等具有明显的调控作用,对抑制肠道炎症、维持肠道稳态、保护肠道健康等同样具有积极的影响。当前对ACE2在肠道健康领域的研究较少。论文就ACE2通过影响肠道微生态对维持肠道稳态作用的研究进行综述,为发掘ACE2的新功能提供参考。 相似文献
15.
急性肾损伤(acute kidney injury, AKI)目前已成为犬最主要的一种肾系疾病,尤其在老年犬中发病率逐年上升。近年来,针刺疗法在宠物临床已得到广泛的应用,并逐渐从治疗瘫痪等外科疾病向治疗宠物内科疾病的方向发展。本研究以腺嘌呤为造模药物建立急性肾损伤犬模型,采用电针疗法进行治疗。研究电针疗法对于犬的肾功能、钙磷代谢、抗氧化能力以及对于NRF2通路相关蛋白的影响,探究电针疗法对犬的急性肾损伤的治疗作用。选取24只3～4 kg健康比格犬,随机分为对照组、造模组、常规治疗组、电针干预组、电针治疗组和联合治疗组。第1～15天为造模期,第16～30天为治疗期。试验结束后检测尿比重、血清中尿素氮(BUN)、肌酐(CREA)、尿酸(UA)、钙(Ca2+)和磷(P3+)的变化;影像诊断X光检测肾变化情况;检测血清及肾组织的超氧化物歧化酶(SOD)和丙二醛(MDA);病理组织学观察各组试验犬肾组织病变的严重程度;采用qRT-PCR和Western blot方法检测各组试验犬肾组织中与抗氧化相关基因和蛋白的表达情况;用免疫组织化学的方法检测肾组织中... 相似文献
16.
King JN Strehlau G Wernsing J Brown SA 《Journal of veterinary pharmacology and therapeutics》2002,25(5):371-378
The effect of renal insufficiency was studied on the pharmacokinetics (PK) and pharmacodynamics (PD) of the angiotensin-converting enzyme (ACE) inhibitor benazepril in cats. The active metabolite of benazepril, benazeprilat, is eliminated principally ( approximately 85%) via biliary excretion in cats. A total of 20 control animals and 32 cats with moderate renal insufficiency induced by partial nephrectomy were used. Assessments were made at steady state after treatment with placebo or benazepril (0.25-2 mg/kg) once daily for a minimum of 10 days. The PK endpoint was the AUC (0-->24 h) of total plasma benazeprilat. The PD endpoints were systolic, diastolic and mean blood pressures (respectively SBP, DBP and MBP) measured by telemetry, and plasma ACE activity, assessed by an ex vivo assay. Renal function was assessed by glomerular filtration rate (GFR), measured by inulin clearance, and plasma creatinine concentrations (1/PCr). As compared with control animals, the renal insufficient cats had a 78% reduction in GFR (0.57 +/- 0.41 mL/min kg), increased plasma creatinine (2.7 +/- 1.0 mg/dL), urea (44.0 +/- 11.9 mg/dL) and ACE activity, and moderately increased blood pressure (SBP 171.8 +/- 5.1 mmHg) (all parameters P < 0.05). Renal insufficient cats receiving benazepril had significantly (P < 0.05) lower SBP, DBP, MBP and ACE, and higher GFR values as compared with placebo-treated animals. There were no significant differences in SBP, DBP, MBP, benazeprilat or ACE values according to the degree of renal insufficiency in cats receiving benazepril. It is concluded that no dose adjustment of benazepril is necessary in cats with moderate renal insufficiency. 相似文献
17.
旨在克隆牛HSL基因的CDS序列,并对该基因进行生物信息学、组织表达谱分析。根据GenBank登录的牛HSL基因序列(NM_001080220)设计1对引物,对牛组织样品中的HSL基因CDS序列进行RT-PCR扩增及序列测定;利用半定量RT-PCR方法检测HSL基因在牛各组织中的表达情况;利用生物信息学软件对其所编码的牛HSL蛋白进行分析。结果显示,获得的牛HSL基因CDS全长为2271bp,编码756个氨基酸,与GenBank登录的牛HSL基因同源性最高,为99.9%。HSL蛋白含有一个HSL-Nsuperfamily保守结构域,无信号肽结构,具有疏水性。半定量RT-PCR显示,HSL基因在检测的心脏、脾脏、肺脏、肾脏、肝脏、大网膜、皮下脂肪、肌肉8种组织中均有表达,其中在大网膜和皮下脂肪中表达量较高,在肾脏、脾脏、肝脏、肺脏、肌肉和心脏中度表达。该试验可为研究牛HSL蛋白的结构和功能提供参考。 相似文献
18.
旨在克隆血管紧张素转换酶2（angiotensin converting enzyme 2,ACE2）基因,分析其结构特性和表达特征。本试验以45和90日龄的健康雪山草鸡为研究对象,利用PCR技术扩增ACE2基因的编码区序列;运用生物信息学方法对其结构特征进行分析;通过TA克隆构建黄羽肉鸡ACE2编码区序列质粒标准品,使用绝对定量方法测定ACE2基因在45和90日龄雪山草鸡公母鸡各组织中的表达特性。结果,克隆得到2 427 bp的编码区序列,共编码808个氨基酸,与白羽肉鸡一致,但是在黄羽肉鸡序列中存在大量突变位点造成编码氨基酸发生变化。同源性比较发现,其与红色原鸡的同源性为99%,其次为绿头鸭88%,与其它动物的同源性为73%~76%。生物信息学分析发现,ACE2属于I型跨膜蛋白,即分泌型蛋白,信号肽位于1~17 aa,含有46个蛋白质磷酸化位点。定量结果发现,小肠中ACE2表达量显著高于其它组织。法氏囊、气管、胆囊和脾中ACE2表达量较低,脾中表达量最低。母鸡空肠、盲肠、脾和肺中ACE2基因表达量显著高于公鸡。公鸡肾和心中ACE2基因表达量显著高于母鸡。90日龄雪山草鸡的十二指肠、空肠、肺、脾和回肠组织ACE2基因表达量均显著低于45日龄雪山草鸡,在法氏囊和肾组织中结果呈相反趋势。通过克隆获得了雪山草鸡ACE2基因编码区序列,生物信息学分析发现,雪山草鸡ACE2基因在家禽中较为保守;ACE2基因表达规律呈现出在雪山草鸡肠道组织中高于其他组织,母鸡中高于公鸡,随着时间表达水平呈下降趋势。研究结果为开展ACE2基因在黄羽肉鸡肠道中的功能研究奠定了理论基础。 相似文献
19.
旨在探讨牦牛（Bos grunniens）肾集合系统的解剖结构。运用丙烯腈-丁二烯-苯乙烯（acrylonitrile butadiene styrene，ABS）铸型技术，制作24个牦牛肾集合系统标本，通过观察标本，记录肾集合系统的解剖特点。结果显示：肾盂在所有标本中都存在，根据输尿管近端有无扩张分为两种类型：扩张型肾盂和未扩张型肾盂，后者更多见（58.3%）。集合系统中有前、后两个肾大盏，肾小盏通过漏斗状结构与肾大盏相连，一个漏斗状结构连接1～5个肾小盏。牦牛肾集合系统前后区域肾小盏数量差异显著（P<0.01），其中，后部肾小盏数量比前部多。根据肾前后区域和中间区域尿液收集引流特点，将牦牛肾集合系统分为两种类型：A型和B型。肾中间区域肾实质尿液的收集和引流依赖前、后肾盏组的为A型，独立于前、后肾盏组的为B型，在牦牛中，A型是常见类型，占70.8%，其次是B型，占29.2%。牦牛肾集合系统由1个肾盂、2个肾大盏和多个（11～21）肾小盏构成，根据尿液引流特点分为A、B两种类型，其中A型较常见。 相似文献
20.
This work aims to study the anatomical structure of the yak kidney collection system. Twenty-four collecting system casting specimens, which were made by ABS (acrylonitrile butadiene styrene), were observed to study its anatomical structure. The results showed that the renal pelvis was present in all casts and is classified into 2 types, dilated or nondilated (58.3%), with or without the dilated origin of the ureter. All casts had 2 major renal calyces toward the cranial and the caudal poles. Minor renal calyces were connected to the major renal calyces by an infundibulum, which drained 1 to 5 minor renal calyces. The number of minor renal calyces in the caudal region was significantly more than that in cranial (P<0.01). The yak kidney collection system was classified into 2 types according to the character of drainage. The type A, which was composed of the collection system that presented two major caliceal groups and mid-zone drainage was dependent on these major groups, and type B, which mid-zone drainage was independent of the polar caliceal groups, the type A (70.8%) was more common than type B (29.2%) in the yak. The collection system of yak kidney was composed of 1 renal pelvis, 2 major renal calyces, and multiple (11-21) minor renal calyces. According to the characteristics of drainage, it was divided into 2 types, A and B, of which type A is more common. 相似文献