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1.
The purpose of this work was to study whether the bovine leukocyte adhesion deficiency (BLAD) allele is present in native cattle breeds and the Holstein breed in Turkey. Blood samples were obtained from 120 Holstein, 20 Brown Swiss, 20 Anatolian Black, 20 Turkish Grey, 20 South Anatolian Red and 20 East Anatolian Red cattle. The isolated DNA materials were multiplied in PCR using the primer developed by Kriegesmann et al. (1997). In order to determine the area of mutation in PCR products, the PCR products were digested with TaqI endonuclease enzyme. The resulting fragments were analysed on 2% agarose gel for the absence of a TaqI restriction site. It was found that two of the Holstein cattle (a bull and a cow) were heterozygote BLAD carriers. There was no homozygote BLAD animal. The BLAD allele was not found in the other breeds used in the study. The mutant BLAD allele frequency in the 120 Holstein cattle calculations was 0.0084.  相似文献   

2.
根据GenBank登录的鹅细小病毒(GPV)和番鸭细小病毒(MDPV)非结构蛋白(NS)基因特征,本研究设计1对特异性引物对GPV和MDPV基因组DNA进行PCR扩增,目的片段大小均为810 bp,并对PCR产物进行切胶回收。用EcoRⅠ酶对GPV和MDPV特异性胶回收产物进行酶切鉴定,结果显示MDPV经EcoRⅠ酶切后琼脂糖凝胶电泳检测片段为2段,大小为530和280 bp;而GPV经EcoRⅠ酶切后琼脂糖凝胶电泳检测片段大小不变。本研究建立了一种快速区别GPV和MDPV感染的检测方法,可对番鸭感染水禽细小病毒的情况进行快速鉴别诊断。  相似文献   

3.
Hemophilia A in three unrelated cats   总被引:1,自引:0,他引:1  
Factor VIII deficiency, consistent with hemophilia A, was detected in 3 unrelated male domestic short-haired cats. Clinical history of each case differed and illustrated the spectrum of clinical manifestations of this disease in cats. Clinical expression of feline hemophilia A was similar to that of man and other animals, in which the degree of factor VIII deficiency tends to determine the severity and frequency of bleeding. All 3 cats had severe protracted bleeding after minor surgical procedures. Hemorrhage was eventually controlled in each case by transfusions of fresh whole blood.  相似文献   

4.
AIM: To investigate cases of coat-colour dilution and hypotrichosis in a group of Hereford x Friesian crossbred calves, and to define the underlying molecular genetics of the disorder. METHODS: The investigation was predicated on the hypothesis that this disorder was similar to a known dominantly inherited disorder of calves of black breeds crossed with Simmental cattle, for which there were candidate gene mutations. Sequence analyses of PCR amplicons from exon 1 and exon 11 of the premelanosome protein 17 gene (PMel17) were carried out. Restriction enzyme digestions of amplicons were followed using electrophoresis of digested fragments. RESULTS: It was shown that an affected calf and its Hereford sire were heterozygous for a three-base deletion in exon 1 of the PMel17 gene. These two animals were also heterozygous for a second mutation in exon 11 of the PMel17 gene. Four other related animals were likewise heterozygous for both mutations in the sire's herd of origin. CONCLUSIONS: Coat-colour dilution and hypotrichosis in Hereford crossbred calves in New Zealand is the same genetic disorder as that previously described in Simmental crossbred calves, and is linked to mutations in the PMel17 gene.  相似文献   

5.
Jergensen, C. B., J. S. Agerholm, J. Pedersen and P. D. Thomsen: Bovine leukocyte adhesion deficiency in Danish Holstein-Friesian Cattle I. PCR screening and allele frequency estimation. Acta vet. Scand. 1993,34,231-236.–A screening program for bovine leukocyte adhesion deficiency (BLAD) in Danish Holstein-Friesian cattle has been initiated. During the first months 1611 animals were tested by a PCR based assay. Of these animals 1256, 346, and 8 were assigned normal, BLAD carriers, and BLAD affected animals, respectively One bull, born as a co-twin, showed weak reaction for the BLAD allele on DNA isolated from leukocytes, but a normal genotype on DNA isolated from semen. Chromosome analysis showed that this bull was a blood chimaera. Estimation of the BLAD allele frequency upon the PCR test results showed that around 450 Danish calves born in 1991 might have been affected with the recessive disorder.  相似文献   

6.
A coagulopathy with subcutaneous bleeding and muscular or peritracheal/periesophageal bleeding occurred in two male Japanese Brown calves of the same dam. One of the affected calves died three days after the onset of bleeding and the other survived normally until being slaughtered despite once suffering from subcutaneous hematoma. Hemostatic tests of the latter case showed prolonged activated partial thromboplastin time (APTT), and severely reduced factor VIII activity. In addition, von Willebrand factor activity, determined by the human platelet aggregation test, was within the normal range; therefore, the calf was diagnosed with hemophilia A. These are the first bovine cases of hemophilia A definitely diagnosed clinicopathologically.  相似文献   

7.
Hemophilia A is an X‐chromosome‐linked disorder caused by a deficiency in factor VIII (FVIII). Although foals have been diagnosed with hemophilia A based on deficiency in FVIII activity, causative gene mutations have not been identified. The genomic DNA and cDNA encoding FVIII of a Tennesee Walking Horse colt affected with hemophilia A and the genomic DNA of his dam and a normal unrelated horse were analyzed with no splice site or coding sequence abnormalities identified in any of the horses. Polymerase chain reactions (PCR) were then performed on hepatic cDNA from the affected colt and an unrelated normal horse, and no product was obtained for the sequence between and including exon 1 and exon 2 in the affected colt. Based on these results, suspected mutations were identified in the noncoding region of FVIII (intron 1), and genomic sequencing of intron 1 in the dam and the affected colt suggested maternal inheritance.  相似文献   

8.
Polymorphisms in the prion protein gene ( PRNP ) are known to be associated with transmissible spongiform encephalopathies in human, sheep and goats. There is tentative association between PRNP promoter polymorphism and bovine spongiform encephalopathy (BSE) susceptibility in cattle. In this study, we genotyped for six bovine PRNP polymorphic sites including a 23-bp indel in the promoter, a 12-bp indel in the intron 1, two nonsynonymous single nucleotide polymorphisms (SNPs), octapeptide repeats in the coding region and a 14-bp indel in the 3'-untranslated region in 178 animals representing Japanese Brown, Kuchinoshima feral, Mishima, Japanese Shorthorn and Holstein. In 64 Japanese Brown cattle, three indel sites were polymorphic. All of the six sites were monomorphic in Kuchinoshima. The 23-bp and 12-bp indel sites were polymorphic in Mishima cattle. The 23-bp and 14-bp indel sites were polymorphic in Japanese Shorthorn cattle. Both SNP sites were monomorphic in all cattle examined in this study. At the 23-bp indel site, the genotype frequencies of Japanese Brown and Holstein breeds were similar to that of BSE affected cattle. We estimated 12 different haplotypes from these genotypic data. A '23-12-K6S14+' haplotype was the major haplotype in all populations, whose frequencies ranged from 0.50 to 1.00.  相似文献   

9.
Forelimb‐girdle muscular anomaly is a hereditary disorder of Japanese Black cattle characterized by tremors and astasia caused by hypoplasia of the forelimb‐girdle muscles. The locus responsible for this disorder has been mapped on a middle region of bovine chromosome 26. In this study, we applied marker‐assisted selection to identify the carriers of this disorder. Four microsatellite markers, DIK4440, BM4505, MOK2602 and IDVGA‐59, linked to the disorder locus were genotyped in 37 unaffected offspring of a carrier sire. Transmission of the mutant or wild‐type allele of the disorder locus of the sire to the 37 offspring was determined by examining the haplotypes of these markers. The results showed that nine and 18 of the 37 animals possessed the paternally transmitted mutant and wild‐type alleles, respectively, and therefore, the nine animals with the mutant allele were identified as carriers. We concluded that the marker‐assisted selection using these four markers can be applied for the identification of the carriers of forelimb‐girdle muscular anomaly of Japanese Black cattle.  相似文献   

10.
本研究根据Genbank中牛肌肉生成抑制素基因序列设计引物,在设计的引物两端分别加上限制性内切酶BarnHI和EcoRI的识别位点序列。利用RT—PCR技术从西门塔尔牛肌肉组织的总RNA中扩增出MSTN基因的CDNA序列,扩增出1125bp片段,该片段与pMD18-T载体连接,转化JM109感受态细胞,所得阳性克隆进行酶切和PCR鉴定,并进行了测序分析,得到的克隆序列与设计的序列基本一致,表明成功地克隆了西门塔尔牛的肌肉生成抑制素基因的蛋白编码序列。  相似文献   

11.

Background

Bovine leukocyte adhesion deficiency (BLAD), deficiency of uridine monophosphate synthase (DUMPS), complex vertebral malformation (CVM), bovine citrullinaemia (BC) and factor XI deficiency (FXID) are autosomal recessive hereditary disorders, which have had significant economic impact on dairy cattle breeding worldwide. In this study, 350 Holstein cows reared in Turkey were screened for BLAD, DUMPS, CVM, BC and FXID genotypes to obtain an indication on the importance of these defects in Turkish Holsteins.

Methods

Genomic DNA was obtained from blood and the amplicons of BLAD, DUMPS, CVM, BC and FXID were obtained by using PCR. PCR products were digested with TaqI, AvaI and AvaII restriction enzymes for BLAD, DUMPS, and BC, respectively. These digested products and PCR product of FXID were analyzed by agarose gel electrophoresis stained with ethidium bromide. CVM genotypes were detected by DNA sequencing. Additionally, all genotypes were confirmed by DNA sequencing to determine whether there was a mutant allele or not.

Results

Fourteen BLAD, twelve CVM and four FXID carriers were found among the 350 Holstein cows examined, while carriers of DUMPS and BC were not detected. The mutant allele frequencies were calculated as 0.02, 0.017, and 0.006 for BLAD, CVM and FXID, respectively with corresponding carrier prevalence of 4.0% (BLAD), 3.4% (CVM) and 1.2% (FXID).

Conclusion

This study demonstrates that carriers of BLAD, CVM and FXID are present in the Turkish Holstein population, although at a low frequency. The actual number of clinical cases is unknown, but sporadic cases may appear. As artificial insemination is widely used in dairy cattle breeding, carriers of BLAD, CVM and FXID are likely present within the population of breeding sires. It is recommended to screen breeding sires for these defective genes in order to avoid an unwanted spread within the population.  相似文献   

12.
The congenital bleeding disorders of domestic animals usually mimic closely the same disorders in man and are inherited in a similar fashion. Classical haemophilia (haemophilia A, factor VIII deficiency) and haemophilia B (factor IX deficiency) occur in both dogs and cats and are sex-linked conditions. Affected animals are almost always male and heterozygote females are asymptomatic carriers. Offspring of a carrier female have a 50 per cent chance of inheriting the defective X-chromosome and, on average, half of the sons will be affected and half of the daughters will be carriers. Diagnosis of affected animals is confirmed by specific factor assay. Carriers may be identified with a statistical certainty of greater than 80 per cent. Canine von Willebrand's disease is a less severe disorder due to a defect of platelet adhesion. It is an autosomal trait, inherited in most breeds in an incompletely dominant fashion.  相似文献   

13.
Objective – To describe the clinical course of a dog with hemophilia A and circulating factor VIII inhibitors complicating the treatment of postoperative hemorrhage.
Case Summary – A 7-year-old castrated male Japanese Chin with hemophilia A, weighing 6 kg, was presented for dental cleaning, polishing, and extractions. Despite presurgical administration of cryoprecipitate, continuous oral bleeding occurred. Circulating factor VIII inhibitors were detected, and the postoperative hemorrhage was subsequently managed with extensive and prolonged blood component transfusion. The dog was discharged after a full clinical recovery.
New or Unique Information Provided – This case report describes the clinical consequences and successful treatment of postoperative hemorrhage in a dog with hemophilia A and circulating factor VIII inhibitors. A relevant discussion of the management of human patients with circulating factor VIII inhibitors is included.  相似文献   

14.
[目的]为了探究广西南宁市肉牛的父系遗传背景与遗传组成。[方法]利用PCR扩增、限制性酶酶切和生物信息学方法,对南宁屠宰场的73头肉牛Y染色体USP9Y基因的遗传多态性进行分析。[结果]发现73头公牛USP9Y基因的PCR产物具有多态性,2头牛显示471 bp带型,71头牛显示552 bp带型。在71个552 bp带型中,有28个可以被SspI酶切成2条带(338 bp和215 bp),表明这28头牛为Y3单倍型组(38.36%),而其余43个不能被SspI酶切,表明这43头牛为Y2单倍型组(58.90%)。仅有2头牛的PCR产物为471 bp,表明这2头牛为Y1单倍型组(2.74%)。屠宰牛群的单倍型多样度为0.5122±0.0309,表明屠宰牛群的Y染色体遗传多样度较高。[结论]南宁市屠宰牛群的来源比较复杂,有普通牛(Y1与Y2单倍型组)和瘤牛(Y3单倍型组)2个父系起源。  相似文献   

15.
试验旨在对宁夏六盘山区肉牛杂交改良群体肌肉生长抑制素(myostatin,MSTN)基因的多态性进行研究,为宁夏肉牛杂交改良选育提供理论基础。采用PCR-RFLP对该群体110头个体MSTN基因多态性进行检测。结果扩增出1346 bp的目的片段,其PCR产物被限制性内切酶DraⅠ消化后表现出多态性,共检测到2种基因型:AT和TT,其中AT基因型频率为0.0545,TT基因型频率为0.9455;T为优势等位基因,TT基因型为优势等位基因。因此,宁夏六盘山区的肉牛杂交改良群体MSTN基因具有多态性。  相似文献   

16.
试验通过对宁夏六盘山区肉牛杂交改良群体生长激素受体(growth hormone receptor,GHR)基因的多态性研究,为宁夏肉牛杂交改良选育提供理论基础。采用PCR-RFLP对该群体70个个体GHR基因多态性进行检测,结果扩增出338 bp的目的片段,PCR产物被限制性内切酶Alu Ⅰ消化后表现出多态性,共检测到2种基因型:AA、AB,其中A为优势等位基因,AA基因型为优势基因型。  相似文献   

17.
孙彦  郭校民  周禾 《草业科学》2012,29(4):544-548

摘要:根据表达载体PBI 121特性及酶切位点设计合适的引物,由表达引物通过PCR技术从含有白颖苔草(Carex rigescens)CrHsf全长cDNA的克隆载体的大肠杆菌上扩增出带有特定酶切位点的CrHsf完整开放阅读框。再对载体和目的片段进行酶切处理,处理后将正确的目的基因片段亚克隆至PBI 121植物表达载体。通过PCR及酶切鉴定,结果证明,目的基因片段已被正确克隆到表达载体上,载体构建成功。  相似文献   

18.
本试验旨在对宁夏六盘山区肉牛杂交改良群体垂体特异性转录因子1(Pit-1)基因的多态性进行研究,为其杂交改良提供理论基础。采用聚合酶链式反应—限制性片段长度多态性方法(PCR-RFLP),对该群体101个个体Pit-1基因多态性进行检测。结果表明,扩增出的451 bp片段为目的基因片段,其PCR产物经限制性内切酶HinfⅠ消化后表现出多态性,共检测到AA、BB和AB 3种基因型。其中AA基因型频率6.93%(7个);AB基因型频率34.65%(35个);BB型基因型频率58.42%(59个),等位基因A频率24.26%;等位基因B频率75.74%。因此,宁夏六盘山地区的肉牛杂交改良群体Pit-1基因具有多态性。  相似文献   

19.
OBJECTIVE: To develop a molecular genetic test to detect the mutant skeletal muscle chloride channel (CIC-1) allele that causes myotonia congenita in Miniature Schnauzers and to analyze the relationship of affected and carrier dogs. ANIMALS: 372 Miniature Schnauzers from the United States, Canada, Australia, and Europe that were tested between March 2000 and October 2001. PROCEDURE: The sequence surrounding the mutation in the CIC-1 allele was amplified by use of a unique pair of primers. Polymerase chain reaction (PCR) products were digested with the restriction enzyme Hpy CH4 III and separated on a 6% polyacrylamide gel. Pedigrees from all available carrier and affected dogs were analyzed, and a composite pedigree was established. RESULTS: Enzyme digestion of PCR products of the normal CIC-1 allele resulted in 3 fragments of 175, 135, and 30 bp, whereas PCR products of the mutant allele resulted in fragments of only 175 and 165 bp. Of the 372 Miniature Schnauzers, 292 (78.5%) were normal, 76 (20.4%) were carriers, and 4 (1.1%) were affected (myotonic) dogs. Frequency of the mutant allele was 0.113. Pedigree analysis revealed that a popular sire, documented to be a carrier, was a common ancestor of all carriers and affected dogs. CONCLUSIONS AND CLINICAL RELEVANCE: A PCR-based enzyme digestion DNA test was developed. The mutant allele for this disease is frequent in Miniature Schnauzers that are related to a common carrier ancestor. Breeding dogs should be tested by this specific DNA test to help limit the spread of this deleterious mutation.  相似文献   

20.
Renal tubular dysplasia is an autosomal recessively inherited disorder in Japanese black cattle that is due to deletion mutations in the claudin-16 gene and causes chronic renal failure and death of affected animals. Here, we report a multiplex-PCR procedure to determine the genotype for claudin-16 deficiency in preimplantation embryos. The presence or absence of the wild-type and mutant allele(s) was precisely detected with the multiplex-PCR using as little as 5 pg of genomic DNA from leukocytes. When biopsied embryo cells were examined for claudin-16 deficiency, 97.2% of genotypes were consistent with the PCR results obtained for the corresponding embryos. In addition, sexing of embryos by PCR was performed using an aliquot of DNA extracted from biopsied embryo cells, and determination of claudin-16 genotype and sex was successfully achieved with an efficiency of 91.7% for claudin-16 genotyping and 83.3% for sexing. The production of a 100-day fetus that was male and homozygous for claudin-16 deficiency, as expected from the analysis of biopsied embryo cells, gave evidence of the reliability and applicability of this procedure for preventing the transmission of this disease and for enabling advances in animal breeding.  相似文献   

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