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禽传染性支气管炎病毒免疫原基因组的RT—PCR研究 总被引:9,自引:0,他引:9
本文报道了用聚合酶链反应获取IBV M41株和广东肾变栋D41免疫基因的研究情况,所用引物为IBV Beaudette株S1基因两侧的对应序列,跨幅为1.7kb;DNA模板为IBVM41和D41株经病毒RNA提取后反转录而得;2株病毒所获的PCR产物与预期一致。 相似文献
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选择S1基因做为目的基因,运用RT-PCR技术分别扩增12株以非免疫鸡胚繁殖并通过超速离心浓缩的鸡胚尿囊液中的IBV,所用引物为IBVBeaudette株S1基因两侧的对应序列,跨幅为1.72kb。结果6株IBV毒株(SAIB3、F、D41、M41、H52、Holte)扩增出了长约1.7kb的S1基因片段,而另6株IBV毒株虽经4次RT-PCR重复后也显阴性。用HaeⅢ内切酶对6个毒株的RT-PCR产物进行酶切,结果显示M41、H52、D413个IBV毒株的S1基因包含两个HaeⅢ位点,F株与Holte株S1基因包含1个HaeⅢ位点,而SAIB3株S1基因则已丢失HaeⅢ位点。实验结果表明:运用RT-PCR技术检测IBV时,由于S1基因的核酸序列具有较高的变异率,因此S1基因不宜做为目的基因。 相似文献
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以IBVBeaudette株S1基因两仙21个碱基的对应序列来合成引物,并在两引物Oligo5′,Oligo3′的5′端分别加有HindⅢBamHI酶切位点,对IBV标准毒株M41,广东地方肾病变形致弱株D41作RT-PCR(反转录-聚合酶链反应)得到了1720bp预计大小的DNA片段,又经M41株PCR产物的HaeⅡ酶切分析,表明此DNA片段即为IBVS1基因片段,并用此对引物,以Oligo3′ 相似文献
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传染性支气近炎病毒类M41地方分离株S1基因克隆及高变区序列分析 总被引:4,自引:0,他引:4
传染性支的管炎病毒(IBV)分离株QD经RT-PCR扩增出约1.7kbS1糖蛋白基因cDNA,修饰后插入pUC18SmalⅠ/EcoRI位点构成质粒pUCQDS1。对克隆的S1基因5′端高变区序列测定显示,起始密码上游60碱基和下游380碱基中与参考株M41S1基因序列仅有1个碱基的差异,表明QD为一类M41分离株。 相似文献
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以人工合成的针对血清I号马立克氏病病毒(MDV1)A抗原基因(gA)部分序列的两端寡聚核苷酸为聚合酶链反应(PCR)的引物,分别对马立克氏病病毒血清I型(京-1株,MD11/75C株);2型(SB-1株);3型(火鸡疱疹病毒的FC126株)毒株和GA株BamHI B片段的PACY184克隆重组质粒DNA进行基因扩增反应及其敏感性试验。结果表明,该引物不仅对MDV1具有较强的特异性,而且由此引物引导 相似文献
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传染性支气管炎病毒中国地方分离株RFLP基因分型的研究 总被引:5,自引:1,他引:4
为初步确定我国不同地区流行的传染性支气管炎病毒(IBV)的基因分型,对分离自国内8个不同地域疫区的IBVQD、GZ、ZZ、TJ、DL、YC、JS1和JS2及参考株M41、H52和T的S1基因RT-PCR扩增cDNA进行HaeⅢ的RFLP分析。结果,QD与MD41、H52同属Massachussete基因型,GZ、ZZ、YC与T的基因型相同,DL、JS1和JS2则表现为各自独立的基因型,而TJ则为DL和T2种基因型毒株的混合感染,表明我国的广大地域内存在着Mass基因型、T基因型和可能的变异株IBV的流行。 相似文献
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用单抗介导的荧光抗体技术检测鸡尿囊细胞内传染性支气管炎病毒的研究 总被引:2,自引:0,他引:2
用传染性支气管炎病毒(IBV)H120、H52、M41、ARK、澳大利亚T株、野外分离株RS、RY及鸡新城疫病毒(NDV)、鸡传染性法氏囊病毒(IBDV),鸡传染性喉气管炎病毒(ILTV)、减蛋综合症病毒(EDSV)等分别接种9日龄SPF鸡胚,37℃孵育48小时后,将其尿囊液离心,沉淀用PBS悬浮,涂片,用抗IBV单抗MC作一抗、进行间接荧光抗体检测。结果:H12O、H52、M41“ARK、T株及RS、RY均显阳性,而NDV、IBDV、ILTV、EDSV均为阴性。结果表明、用此法检测鸡胚尿囊液中IBV.具有快速、敏感、特异的优点。 相似文献
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多聚酶链反应区分血清Ⅰ型MDV强弱毒株 总被引:1,自引:0,他引:1
选用血清Ⅰ型马立克氏病病毒(MDV_1)基因组中位于132bp正向重复序列两侧的2段寡聚核苷酸为多聚酶链反应(PCR)的引物,分别对Ⅰ型MDV强毒株(京-1株)和弱毒株(Md11/75C株)的核酸进行基因扩增。结果显示:该弱毒株核酸基因中含6个或更多个拷贝数的132bp正向重复序列,而强毒株核酸基因中仅含2个,相同引物对Ⅱ型、Ⅲ型MDV(SB-1株,Fc126株)和禽网状内皮组织增生病病毒(REV)核酸作PCR扩增,未检出任何核酸片段。 相似文献
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应用RT—PCR技术扩增禽传染性支气管炎病毒S1基因的研究 总被引:2,自引:0,他引:2
选择S1基因做为目的基因,运用RT-PCR技术分别扩增12株以非免疫鸡胚繁殖并通过超速离心浓缩的鸡胚尿囊液中的IBV,所用引物为IBV Beaudette株S1基因两侧的对应序列,跨幅为1.72kb。结果6株IBV毒株(SAIB3、F、D41、H52、Holte)扩增出了长约1.7kb的S1基因片段,而另6株IBV毒株虽经4次RT-PCR重复后也显阴性。用HaeⅢ内切酶对6个毒株的RT-PCR产物 相似文献
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魏静 《四川畜牧兽医学院学报》2009,(4):28-32
在现代法律秩序中,商会自治规范是制定法的基础和必要的补充,甚至在某些方面替代了制定法;商会自治规范主要包括商会组织规范、行为规范、惩罚规范以及争端解决规范等;其效力仅及于其内部成员;商会自治规范和制定法之间存在冲突,但也存在整合的基础。 相似文献
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本文概述了猪的毛色类型、猪的毛色遗传模式,着重综述了猪毛色基因分子基础的研究进展,指出存在问题并就未来发展方向做了思考。 相似文献
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以国际标准强毒R株人工感染非免疫产蛋鸡,定时扑杀,分别从鼻窦、眶下孔、气管、肺、气囊、卵巢和输卵管分离MG,并收集感染鸡所产蛋分离MG。结果表明,人工感染48小时后上、下呼吸道及肺已被全面感染,96小时气囊已被感染,120小时输卵管已能分离到MG,卵巢始终分离不到MG。人工感染鸡自144小时便能在其所产蛋中分离出MG。药物治疗能在72小时内消除感染,油乳剂苗则需24天后逐渐降低蛋内MG分离率,药物卵内注射、种蛋药浴、高温处理均能杀死卵内MG,但以研制的种蛋浸泡剂药浴效果为最好。 相似文献
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REASONS FOR PERFORMING STUDY: Centesis of the bicipital bursa using an 8.9 cm long spinal needle has been reported but the alternative of employing a 3.8 cm long hypodermic needle requires validation. OBJECTIVE: To compare the efficacy of 2 different methods of centesis of the bicipital bursa and to evaluate the usefulness of ultrasonographic imaging to determine the location of solution administered when centesis of the bursa is attempted. METHODS: For Trial 1, 6 clinicians, who had no previous experience of centesis of the bicipital bursa, attempted to inject a solution composed of an aqueous radiopaque contrast medium and physiological saline solution (PSS) into the bicipital bursae of 2/12 horses using the previously described distal approach to inject one bursa and a proximal approach to inject the contralateral bursa. The bicipital tendon and bursa were examined ultrasonographically before and after injection; and both shoulders were examined radiographically to identify the location of the medium. In Trial 2, another 6 clinicians, also with no previous experience of centesis, repeated Trial 1, using 6 horses, but the radiopaque contrast medium was mixed with air instead of PSS. RESULTS: Accuracy of centesis using the proximal approach was 39% and that of the distal approach 28%. Ultrasonographic examination of the shoulder allowed the location of solution and air to be accurately predicted in all 12 shoulders examined. CONCLUSIONS: Clinicians who have had no previous experience performing centesis of the bicipital bursa are unlikely to be successful in centesis using either approach. Radiographic examination after injecting a radiopaque contrast medium may be necessary to assess the success of centesis especially if bursal fluid is not obtained during centesis. Injecting air along with the radiopaque contrast medium provides more accurate ultrasonographic confirmation of centesis and better radiographic definition than does injection without air. 相似文献
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Effects of size of ingestively masticated fragments of plant tissues on kinetics of digestion of NDF
Ingestively masticated fragments were collected and sized via sieving. Different sizes of esophageal masticate and ruminal digesta fragments, and ground fragments of larger masticated pieces were incubated in vitro, and undigested NDF remaining at intervals of up to 168 h of incubation was determined. The ruminal age-dependent time delay (tau) for onset of digestion of NDF was positively correlated (P < 0.004) with the mean sieve aperture estimated to retain 50% of the fragments between successive sieve apertures (MRA). Degradation rate of potentially degradable NDF (PDF) and level of indigestible NDF were not related (P > 0.10) to MRA of masticated and ground fragments. Estimates of tau were positively related to MRA, with slopes of bermudagrass < corn silage < ruminal fragments of corn silage. It was concluded that fragment size-, and consequently, ruminal age-dependent onset of PDF degradation of a mixture of different fragment sizes results in an age-dependent rate of degradation of the more rapidly degrading of two subentities of PDF. Models are proposed that assume a tau before onset of simultaneous degradation of PDF from two pools characterized as having gamma-modeled age-dependency and age-constant rates. The ruminal age-dependent pool seems to be associated with the faster-degrading pool, and its rate parameter increases with range in MRA in the population of fragments. Conceptually, the ruminal age-dependent rate parameter for PDF degradation seems to represent a composite of several effects: 1) effects of the size-dependent tau; 2) range in MRA of the population of ingestively masticated fragments; and 3) subentities of PDF that degrade via more rapid age-dependent rates compared with subentities of PDF that degrade via age-constant rates. The estimated fractional rates of ruminative comminution of ingestively masticated fragments (0.060 to 0.075/h) were of a magnitude similar to the mean fractional rates of PDF digestion (0.030 to 0.085/h), which implies that ruminative comminution may be first-limiting to fractional rate of PDF digestion. The in vivo roles of ingestive and ruminative mastication of fragments on PDF degradation must be considered in any kinetic system for estimating PDF digestion in the rumen. These results and others in the literature suggest that the rate of surface area exposure rather than intrinsic chemical attributes of PDF may be first-limiting to degradation rate of PDF in vivo. 相似文献