摘要:

SummaryA method for genotyping K‐casein (A, B, E), β‐casein (A1,A2,A3, A5,B) and β‐lactoglobulin (A, B) simultaneously by the use of allele discrimination by primer length combined with automated detection of fragments with a sequencing instrument is described. Seven different mutations within the milk protein genes were analysed in order to distinguish between the alleles examined. The samples were amplified in two separate multiplex polymerase chain reactions (PCRs), which were then pooled and separated according to size in a single lane on the gel. By using stringent PCR conditions, we have been able to achieve allele‐specific amplifications and minimize amplification of mismatched primer for all seven

ISSN:0268-9146    

DOI:10.1111/j.1365-2052.1995.tb02635.x

出版商:Blackwell Publishing Ltd    

年代:1995

数据来源: WILEY

摘要:

SummaryMicrosatellite polymorphisms are finding increasing use in genetics. In addition to the random isolation of microsatellite markers, such markers can also be developed from sequences already present in public domain databases. An advantage of public domain databases is that these microsatellites are known to be located within or close to identified functional genes. In this study the GenBank and EMBL databases were screened for microsatellite markers and primers were defined for amplification. Subsequently, these markers were tested on a panel of five different birds from layer and broiler stocks and on the international reference families: the East Lansing reference family and the Compton reference family. Of the 33 loci tested, 25 were polymorphic on the test panel and from these 25, 14 were polymorphic in one or both reference families. Twelve of the 14 loci that could be mapped fell into previously defined linkage groups. The other two markers were not linked. Because three of the loci had previously been mapped to specific chromosomes byin situhybridization, linkage groups E6 and C3 could be assigned to chromosome 6, E5 and C17 to chromosome 4 and E21 to one of the microchromosomes.

ISSN:0268-9146    

DOI:10.1111/j.1365-2052.1995.tb02636.x

出版商:Blackwell Publishing Ltd    

年代:1995

数据来源: WILEY

摘要:

SummaryThe fine specificities of two panels of monoclonal antibodies (mAbs) for sheep major histocompatibility complex (MHC) class II molecules were determined using five mouse L‐cell transfectaents, each expressing a defined sheepDQorDRMHC class IIA/Bgene pair. Using the transfectants in an indirect fluorescence antibody assay, previous immunochemical characterization of the mAbs was confirmed for 16 of 23 mAbs tested. The MHC class II subtype specificity (DQorDR) of each mAb was assigned without interference from the products of other expressed class II loci. This allowed the identification of both cross‐locus specificities as well as defining fine specificities of mAbs previously only partially characterized by immunochemical techniq

ISSN:0268-9146    

DOI:10.1111/j.1365-2052.1995.tb02637.x

出版商:Blackwell Publishing Ltd    

年代:1995

数据来源: WILEY

摘要:

SummarySix loci, apoliproprotein B (including Ag(x) antigen), immunoglobulin kappa constant region (IGKC), luteinizing hormone/choriogonadotrophin receptor, avian myelocytomatosis viral related oncogene, neuroblastoma derived, ornithine decarboxylase, and proopiomelanocortin (adrenocorticotropin/beta‐lipotropin) (POMC), were newly assigned to sheep chromosome 3p using a chromosomally characterized minipanel of sheep‐hamster cell hybrids. Isotopicin situhybridization of IGKC to sheep chromosome 3p22–p17 is reported, confirming the cell hybrid assignment. As these loci are all known to map to human chromosome 2p, this study demonstrates that this chromosomal segment is extensively conserved in sheep. Only POMC has been previously assigned to cattle chromosome 11, which is the equivalent of sheep chromosome 3p. Therefore, we predict that the other loci assigned in this study to sheep 3p are likely to be located on cattle 11. The provisional assignment of an additional locus, annexin‐like to sheep chromosome 3p is also r

ISSN:0268-9146    

DOI:10.1111/j.1365-2052.1995.tb02638.x

出版商:Blackwell Publishing Ltd    

年代:1995

数据来源: WILEY

摘要:

SummaryMolecular genotyping of swine major histocompatibility complexSLA‐DQBandSLA‐DRBgenes using polymerase chain reaction (PCR)‐based amplification is described. Locus‐specific oligonucleotide primers were designed for the analysis of expressed SLA genes by reverse transcription‐polymerase chain reaction (RT‐PCR). RT‐PCR products were sequenced, and the information gained was used to design primers for PCR genotyping of the exon 2 (β1) region from genomic DNA templates. A single segregating amplification product was detected for both DQB and DRB in all animals. PCR products were digested with restriction enzymes. SevenSLA‐DQBPCR‐restriction fragment length polymorphism (RFLP) pattern types were observed for bothHaeIII andRsal that defined 14SLA‐DQBalleles. A total of sevenSLA‐DRBPCR‐RFLP pattern types were defined usingMspl (3 RFLP pattern types) andRsal (6 RFLP pattern types). In order to demonstrate their universal utility, the primers were tested on genomic DNA samples from 10 different swine breeds. No breed‐specific alleles were observed. These results show that locus‐specific oligonucleotide primers and RFLP analysis provide a simple and rapid method for genotyping expressedSLA

ISSN:0268-9146    

DOI:10.1111/j.1365-2052.1995.tb02639.x

出版商:Blackwell Publishing Ltd    

年代:1995

数据来源: WILEY

摘要:

SummaryThe 5′ flanking region of the α‐lactalbumin (α‐LA) gene was sequenced for the Duroc, Yorkshire and Meishan breeds of swine to identify potential sequence variants within this regulatory region of the porcine α‐LA gene. The sequenced region of the gene encompasses 391 bp 5′ of the translation start site to 11 bp 3′ of the translation start site. Within this sequence of the porcine α‐LA gene two single‐base pair differences were detected. One variant occurs at position – 178 and the other at position – 235 from the translation start site. Each of the variations can be detected by a restriction fragment length polymorphism within a polymerase chain reaction amplified product. The polymorphisms at the – 178 and – 235 positions appear to be genetically linked in the anim

ISSN:0268-9146    

DOI:10.1111/j.1365-2052.1995.tb02640.x

出版商:Blackwell Publishing Ltd    

年代:1995

数据来源: WILEY

摘要:

SummaryUsing agarose gel isoelectric focusing and immunoblotting with rabbit anti‐rabbit C6, a genetic polymorphism was found in the sixth component of complement (C6) in 18 Asian native breeds or populations and three European breeds of dog. The C6 locus was highly polymorphic. The phenotype distribution data indicated that dog C6 phenotypes were controlled by seven codominant alleles,C6A,C6B,C6C,C6D,C6E,C6FandC6G, at a single autosomal locus. Breed differences were observed among the gene frequencies, especially between Asian and European breeds. Two gene flows from the adjacent areas into Japanese native dogs were postulate

ISSN:0268-9146    

DOI:10.1111/j.1365-2052.1995.tb02641.x

出版商:Blackwell Publishing Ltd    

年代:1995

数据来源: WILEY

摘要:

SummaryForty‐eight primer‐pairs complementary to unique DNA sequences flanking chicken (genusGallus) genomic (TG)nmicrosatellite repeats were previously designed. These primer‐pairs were used in the polymerase chain reaction to amplify turkey (genusMeleagris) genomic DNA loci. Results indicated that the majority (92%) of these primer‐pairs generated amplification products in turkey genomic DNA. Hybridization using end‐labelled (TG)8as a probe showed that, out of 41 primer‐pairs tested, only 14 generated an amplification product that also contained a detectable (TG)nmicrosatellite repeat when turkey DNA was the template. Among 18 primerpairs tested for polymorphism, using three commercial turkey lines, five were found to exhibit length polymorphism, three of which did not contain a detectable TG repeat. Therefore, a significant portion of chicken microsatellite markers can be useful for genomic mapping and linkage analysis in the turkey, reducing the costs involved in producing turkey‐specific microsate

ISSN:0268-9146    

DOI:10.1111/j.1365-2052.1995.tb02642.x

出版商:Blackwell Publishing Ltd    

年代:1995

数据来源: WILEY

摘要:

SummaryWe have used a panel of anti‐major histocompatibility complex (MHC) class II monoclonal antibodies (mAbs) and have assessed their specificity for the products of the individual bovine MHC (BoLA) class II subregions. The mAbs identified two distinct class II molecules by affinity purification and ELISA. Two‐dimensional immunoblotting confirmed these data and NH2‐terminal sequencing of the purified class II α chains of one member of each group identified the subregion specificity of the mAbs. The mAbs VPM36, TH22A and TH81A are specific for BoLA DQ, whereas VPM54, TH14B and J11 are specific for BoLA DR. SW73.2 reacts with both MHC subgroups of all cattle

ISSN:0268-9146    

DOI:10.1111/j.1365-2052.1995.tb02643.x

出版商:Blackwell Publishing Ltd    

年代:1995

数据来源: WILEY

摘要:

SummaryThirty (TG)nmicrosatellite clones were isolated from a pig genomic library, sequenced, and tested for their suitability to detect polymorphism on a panel of animals by means of the polymerase chain reaction. Ten of these clones were developed into suitable markers and subsequently segregation of these markers was determined in the five PiGMaP reference pedigrees. A linkage analysis was performed on these 10 microsatellites together with 365 other loci that have been typed on these reference families. Eight of the microsatellites have been mapped to eight different linkage groups that have been previously assigned to different chromosomes (chromosomes 1, 6, 7, 9, 14, 15, 17 and 18). Of the remaining two markers, one is X‐linked and the other shows no linkage. The number of alleles detected by these microsatellites, in the reference pedigrees, varied from six to sixteen and the heterozygosity varied from 42 to 85% in the 26 unrelated founder animals of these reference pedigree

ISSN:0268-9146    

DOI:10.1111/j.1365-2052.1995.tb02644.x

出版商:Blackwell Publishing Ltd    

年代:1995

数据来源: WILEY