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1.
不同绵羊品种FGF5基因的多态性分析   总被引:10,自引:0,他引:10  
根据人和小鼠成纤维细胞生长因子5(FGFS)基因的同源序列设计引物对绵羊基因组进行PCR扩增,将扩增片段进行克隆和测序,并与人和小鼠的成纤维细胞生长因子5基因序列进行同源性比较,确定扩增的片段为绵羊的FGF5基因片段。采用PCR-SSCP技术分析了FGF5基因外显子在小尾寒羊、湖羊、藏羊、中国美利奴等4个绵羊品种的多态性。结果表明:FGF5基因在P1和P2引物扩增片段中存在PCR-SSCP多态性。经克隆测序分析,位于外显子1的引物1扩增产物存在G→T突变,该突变位点使编码的氨基酸发生A→S的改变;引物2扩增产物发生了碱基序列C→T的突变,这一突变位点没有引起编码氨基酸的改变,属于同义突变。对不同绵羊品种的基因型和基因频率统计结果表明,引物1扩增产物小尾寒羊、湖羊、藏羊以等位基因A为主,而中国美利奴羊则以等位基因B为主,且各群体均处于Hardy-Weinberg平衡;引物2扩增产物小尾寒羊、湖羊、中国美利奴羊均以等位基因E为主,而藏羊的基因型频率与其它品种有显著差异,中国美利奴羊在引物2位点的基因频率处于Hardy-Weinberg不平衡状态。  相似文献   

2.
应用随机扩增多态性DNA(random amplified polymorphic DNA,RAPD) 技术分析河南省4个地方鸡的品系,固始鸡快羽GK系、固始鸡慢羽GM系、矮小型绿壳蛋鸡、黄麻羽丝毛乌骨鸡的遗传变异。用186个随机引物对4个鸡种基因组DNA池扩增筛选,40个引物产生多态性。此40个引物经重新优化反应体系后,对120个个体的基因组DNA 进行RAPD分析。40个引物共产生387种扩增片段,扩增产物片段的长度大小在100~2220 bp范围内。利用电泳分析数据分别计算4个品系群体之间的遗传相似系数和遗传距离指数,结果4个鸡种的亲缘关系与它们的引种情况基本一致。同时也证明了RAPD技术可以作为分子标记,很好地检测鸡种群内的遗传变异及区分不同鸡品系。  相似文献   

3.
研究首先采用RAPD技术,利用混合基因池(DNA pool)法,对滩羊体大品系、普通品系进行DNA多态性分析。从100种具有10个碱基的随机引物中,筛选出84种引物在滩羊群体基因组中共扩增出358条带,其中22种引物扩增产物表现为多态(占22%),且扩增出32条有差异的条带,占总带数的8.94%(32/358);62种引物的扩增产物表现为单态(占62%)。滩羊体大品系的特异性条带5个,而普通品系的特异性条带7个。这些特异标记可以用来鉴定滩羊的2个品系。滩羊体大品系和普通品系间的遗传距离为0.136±0.087,表明2个品系之间的亲缘关系很近。  相似文献   

4.
PCR检测犬腺病毒方法的建立   总被引:1,自引:0,他引:1  
利用人工合成的两条引物,对病犬肝脏和细胞培养物冻融上清中的1、2型犬腺病毒DNA进行多聚酶链式反应(PCR)检测;再将扩增产物进行同位素标记,与纯化病毒DNA进行打点杂交。扩增产物经电泳分析结果表明,其大小与设计的引物间的序列大小基本一致;扩增产物经标记后只与犬腺病毒DNA发生杂交反应,表明其为犬腺病毒特异性核酸片段。  相似文献   

5.
鸭沙门菌的随机扩增多态性DNA分析   总被引:5,自引:0,他引:5  
以5株同一血清型的鸭沙门茵为研究对象,分别提取基因组DNA后,用20条随机引物对其进行RAPD分析。结果,20条随机引物中不能扩出任何条带的引物有2条,能扩出条带但扩增图谱中没有特异性条带出现的引物有10条,能扩出条带且扩增图谱具有多态性的引物有8条。对筛选到的具有多态性的8条引物进行分析,共扩增出46个DNA片段,片段大小为0.2~3.2kb,其中5个菌株共有的片段有13条,显示多态性的片段有33条。  相似文献   

6.
本研究首先采用RAPD(Romdom Amplified Polymorphic DNA)技术,利用混合基因池(DNApool)法,对滩羊体大品系、普通品系进行DNA多态性分析.从100种具有10碱基的随机引物中筛选出84种引物可在滩羊群体基因组中得到扩增,共扩增出358条带,其中22种引物扩增产物表现为多态(占22%),且扩增出32条有差异的条带,占总带数的8,94%(32/358);62种引物的扩增产物表现为单态(占62%);另有16种引物扩增非常微弱,不便分析(占16%).根据Nei氏片段共享度公式计算出滩羊的体大品系和普通品系间的遗传距离为0.14±0.25,表明两品系之间的亲缘关系很近.同时在分析过程中发现滩羊体大品系,普通品系均出现了具有品系特征的特异性条带.滩羊普通品系有7条PS-107-550,PS-510-500,PS-512-450,PS-101-450,PS-512-400,PS-511-400,PS-12-370;体大品系有5条PS-509-875,PS-515-875,PS-106-750,PS-1008-700,PS-1010-370;以上引物可作为区分两品系的标记引物,特征性条带可以作为各自的分子标记来进行品系鉴定.  相似文献   

7.
试验旨在研究法国夏洛莱羊与中国美利奴羊(新疆型)杂交对其杂交后代生产性能的影响。本试验以法国夏洛莱羊为父本、中国美利奴羊(新疆型)为母本进行杂交试验,通过杂交试验初步研究对其杂交1代(F1)、回交1代(B1)体增重及毛品质的影响。结果表明,3月龄时,夏洛莱羊×中国美利奴羊(新疆型)F1代体重分别比B1代(试验2组)和中国美利奴羊(新疆型)提高2.8%(P>0.05)和46.5%(P<0.01);6月龄时,体重分别提高4.0%(P<0.01)和28.6%(P<0.01);3月龄、6月龄时B1代(试验2组)的体重分别比中国美利奴羊(新疆型)(对照组)提高42.5%(P<0.01)、23.7%(P<0.01)。在产毛量方面,夏洛莱羊×中国美利奴羊(新疆型)F1代的产毛量与B1代差异不显著(P>0.05),但与中国美利奴羊(新疆型)相比,夏洛莱羊×中国美利奴羊(新疆型)F1代、B1代的产毛量分别极显著降低了20.4%(P<0.01)和20.1%(P<0.01);从增重和产毛总的经济效益来看,与中国美利奴羊(新疆型)相比,夏洛莱羊×中国美利奴羊(新疆型)F1代和B1代总的经济收益分别提高了14.80%、10.88%。因此,法国夏洛莱羊与中国美利奴羊(新疆型)杂交可用于杂交优势生产羊肉,但不宜用于羊毛的生产。  相似文献   

8.
为了解山羊致病性大肠杆菌广西分离株的分子多态性,应用随机扩增多态性方法(RAPD)对山羊致病性大肠杆菌进行分型研究.从8条随机引物中筛选出4条能在10株大肠杆菌中具有较好多态性扩增的随机引物,4条随机引物共扩增出18条DNA片段,10个菌株无共有带谱,显示出良好的扩增多态性.菌株Nx31与Nx32曾被认为是同一菌株的两次分离,但是在RAPD分析中,两株细菌的带谱存在明显的差别,表明RAPD比传统的血清学分型具有更高的分辨性.  相似文献   

9.
对云南黑山羊 (10♂ ,2♀ )的基因组DNA性别决定基因 (SRY)进行了体外PCR扩增的研究 ,结果表明适当引物可特异性扩增出雄性个体的性别决定基因片段 (大小约为 185bp) ,而对雌性个体则不能扩增出任何片段 ;研究还利用已筛选出的性别决定基因特异性引物对少量的胚胎细胞进行了特异性扩增 ,检测了 2 0枚云南黑山羊的胚胎 ,其中有 5枚胚胎可扩增出约为 185bp的特异性片段。以上结果为山羊胚胎早期性别鉴定奠定了技术基础。  相似文献   

10.
RAPD标记与四川黑山羊表型性状的相关性研究   总被引:4,自引:0,他引:4  
利用16个多态性较强、重复性好的随机引物对四川黑山羊多个种群的成年(1.5~2岁)黑山羊96只个体的血样DNA进行扩增,16个引物共扩增出170条多态的RAPD条带.对给定的标记,将“有带(1)”、“无带(0)”两类个体看作独立样本,采用SPSS软件的Compare Means中的Independent-samples T Test来比较两样本各数量性状平均值的差异.并根据两独立样本间个体表型值的分布筛选与性状真实关联的分子标记.结果表明,在黑山羊群体中共筛选到5个分子标记分别与不同性状相关,其遗传基础可能是该标记与控制性状的QTL或主基因连锁.  相似文献   

11.
Mycobacterium avium is an important veterinary pathogen causing avian tuberculosis in birds. The aim of the study was to evaluate the genetic relatedness in M. avium isolates from deep tissues of farmed lesser white-fronted geese with avian tuberculosis and in samples from the farm environment. The strains were analyzed by two PCR-based typing methods, inverted repeat (IR) typing and random amplified polymorphic DNA (RAPD) analysis. The primers for the inverted repeats of the insertion sequences IS1245 and IS1311 were used in IR typing, and the RAPD analysis was performed with six primers. Seven of the nine avian strains yielded an identical pattern in the IR typing, but they could be divided into two groups in the RAPD analysis. The remaining two bird isolates had an identical IR pattern (IR cluster II) which they shared with two environmental isolates. However, the RAPD analysis revealed that these environmental isolates had a RAPD pattern (RAPD cluster VI) distinct and different from either of the bird isolates (RAPD clusters II and IV). In all, four M. avium strains were verified as being inducers of avian tuberculosis in birds, and all were distinct from the three environmental strains identified. Thus, the results did not confirm the preliminary idea that a single strain had caused the epidemic. The polymorphism among M. avium strains highlighted the great biodiversity among an M. avium population even in a limited environmental setting during a short time span, and indicated the high susceptibility to avian tuberculosis of lesser white-fronted geese.  相似文献   

12.
The usefulness of random amplification of polymorphic DNA (RAPD) analysis for typing Indian strains of M. tuberculosis was investigated. M. tuberculosis H37Rv, M. tuberculosis DT and 42 clinical isolates of M. tuberculosis were subjected to RAPD-PCR using 7 random decamer primers. All 7 primers were found to be differentiated and produced specific RAPD profiles. The polymorphic amplicons served as RAPD markers for M. tuberculosis. The dendrograms, obtained by different primers, showed the discriminatory ability of the primers. RAPD analysis provided a rapid and easy means of identifying polymorphism in M. tuberculosis isolates, and it was found to be a valuable alternative epidemiological tool. In addition, the results of the present study showed heterogeneity in the M. tuberculosis strains in the population studied.  相似文献   

13.
Randomly amplified polymorphic DNA (RAPD) analysis was used to differentiate 7 strains of Mycoplasma gallisepticum. Six commercially available primers or primer combinations were screened for their ability to differentiate vaccine and type strains. Although major and minor bands were produced with each primer, many of the primers were unsuitable for strain differentiation. The use of primer 6 and combined primers 3 and 4 resulted in complementary RAPD banding patterns for each M. gallisepticum strain. Eleven different isolates representing 7 different strains were segregated into 7 different patterns, corresponding to the 7 strains.  相似文献   

14.
为探讨DLX3基因的多态性与绵羊羊毛品质及生长性状的关系,本研究以5个品系的中国美利奴羊(新疆军垦型)为试验材料,采用PCR-RFLP方法开展了DLX3基因3′UTR区4个SNPs的多态性检测。通过卡方检验分析了4个SNPs在各品系绵羊中的等位基因频率,采用连锁不平衡分析构建了这4个SNPs的单倍型,最后将单位点和单倍型分别与绵羊羊毛品质和生长性状进行关联分析。结果表明:4个SNPs在5个品系间的等位基因频率均存在极显著差异(P<0.01);超细毛绵羊品系与其他4个品系间的基因型分布均存在极显著差异(P<0.01),2个多胎品系与其他3个品系间的基因型分布同样存在极显著差异(P<0.01);相关分析显示,这4个SNPs及其单倍型均对羊毛卷曲度有显著影响(P<0.05),而对其他羊毛性状及生长性状无显著影响(P>0.05)。由此可见,中国美利奴羊(新疆军垦型)DLX3基因3′UTR区的多态性与绵羊毛发卷曲度性状显著相关,可以尝试使用这些SNPs开展高品质细毛羊的分子标记辅助选择。  相似文献   

15.
Pasteurella multocida causes various respiratory disease symptoms in pigs, including atrophic rhinitis and pneumonia. In the present study, 69 strains of P. multocida were isolated from 443 pigs with respiratory clinical symptoms at 182 farms located throughout South Korea from 2009 to 2010. A multiplex capsular PCR typing assay revealed that 69 strains of P. multocida isolated in this study had the biosynthetic locus of the capsules of either serogroup A (47 strains, 68.1%) or serogroup D (22 strains, 31.9%). The 22 strains positive for serogroup D-specific primers were divided into four clusters and the 47 strains positive for serogroup A-specific primers were divided into 12 clusters according to the results of Random Amplified Polymorphic DNA (RAPD) analysis. P. multocida strains in the present study were susceptible to most of the antimicrobial agents used. An analysis of antimicrobial resistance and virulence gene pattern combined with RAPD indicated that a certain P. multocida strain appeared to be genetically identical, implying the persistence of the strain within a single farm.  相似文献   

16.
家蚕来源肠球菌DNA多态性的RAPD分析   总被引:2,自引:1,他引:1  
运用随机扩增多态性DNA(randomamplifiedpolymorphicDNA,RAPD)技术,对从家蚕消化道中分离的表现为生理生化特性多样性的14个肠球菌分离菌株和3个标准菌株进行了DNA多态性研究。应用筛选的8条随机引物,共扩增出625个位点,其中997%为多态性位点,表明供试菌株具有丰富的DNA多态性。对扩增结果进行聚类分析,供试菌株分为Entfaecalis、Entfaecium、Entcasseliflavus、Entavium等4个类群,与其数值鉴定结果呈相同趋势。  相似文献   

17.
Increasing use of Mycoplasma gallisepticum (MG) live vaccines has led to a need for the differentiation of MG strains. The MG strains MK-7, MS-16, S6, FS-9 and R strains and the MG live vaccine strain F were compared by random amplification of polymorphic DNA (RAPD) in this study. Using RAPD, different patterns were found among the MG strains. In addition to this, we examined the differentiating potential of polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) primers targeted at the crmA, crmB, crmC, gapA, mgc2 and pvpA genes encoding cytadherence-related surface proteins. These proteins may take part in the pathogenesis of MG-induced disease. Differentiation of strain F is based on the identification of restriction enzyme sites in the PCR amplicons. Using HphI enzyme, crmC PCR amplicons produced different RFLP patterns. Digestion of amplicons of gapA-specific PCR with MboI enzyme also produced distinct patterns. Differences were observed among strains R and F by digestion of mgc2 PCR amplicons with HaelIl and VspI enzymes and digestion of pvpA PCR amplicons with AccI, PvulI and ScrFI endonucleases. This method can be used for the rapid differentiation of vaccine strain from wild strains. Differentiation of MG strains is a great advantage for diagnosticians or practitioners and it is useful for epidemiological studies.  相似文献   

18.
选用100个RAPD引物和95对SSR引物进行PCR扩增,旨在构建42份高粱和苏丹草品种资源及2份国审品种高粱-苏丹草杂交种的DNA指纹图谱。结果表明,从100个RAPD引物中筛选到9个多态性高、重复性好的引物,多态性条带比率为64.06%,利用4个核心RAPD引物可以为每份品种构建1张特定的数字指纹,并通过其中1个引物F-01构建了1张能鉴别2个杂交种的RAPD指纹图谱,不过该图谱不能区别皖草3号与其父本Sa。从95对SSR引物中筛选出多态性丰富的引物73对,多态性条带比率为86.06%,通过3对核心SSR引物就可以构建42份高粱和苏丹草的SSR数字指纹,同时利用其中1对SSR引物txp18,寻找到2个杂交种的互补带,从而构建了2个高粱-苏丹草杂交种的SSR指纹图谱,这张SSR指纹图谱不仅能鉴别皖草2号和3号,还可以把杂交种与其亲本区别开来。  相似文献   

19.
应用PAPD方法对鸡败血霉形体DNA多态性的研究   总被引:2,自引:0,他引:2  
用随机扩增多态性DNA(RAPD)方法,通过筛选的5个随机引物(OPH-02,OPH-05,OPH-07,OPH-13,OPG-16),对14株鸡败血霉形体(MG)国际标准株和国内分离株进行了DNA多态性研究。结果表明,5个引物共产生21种条带,其中有3个条带分别为2个菌株所特有,扩增产物片段的长度在150~4500bp之间,所有菌株均有1条共同条带,以OPH-05扩增产物的多态性最丰富。根据样品DNA所获得的菌株间相似性指数表明,D9603与D9607相似指数最高,S6,K3913和D9604三者相互间相似性指数最低。提示RAPD方法可用于霉形体遗传标记的分析。  相似文献   

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