Mapping of microsatellite markers developed from a flow-sorted swine chromosome 6 library

Grimm, David R.; Goldman, T.; Holley-Shanks, Rhonda R.; Buoen, L. C.; Mendiola, John R.; Schook, Lawrence B.; Louis, Charles F.; Rohrer, G. A.; Lunney, Joan K.

doi:10.1007/s003359900388

Cited by 9 publications

(8 citation statements)

References 37 publications

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“…As the focus of gene mapping experiments in farm animals necessarily shifts towards targeted marker development, microdissection is becoming an increasingly important technique. It allows the precise collection of subchromosomal segments, which offers an increase in resolution over production of libraries enriched for whole chromosomes, for example by flow-sorting (GRIMM et al 1997). The scraped products of microdissection can be used for cDNA capture experiments and the identification of genes within the region of interest (BROOKES et al 1995) or, as demonstrated here, for production of highly specific chromosome paints and construction of remarker.…”

Section: Discussionmentioning

confidence: 99%

Microdissection of Pig Chromosomes: Dissection of Whole Chromosomes, Arms and Bands for Construction of Paints and Libraries

et al. 2004

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Chromosome microdissection is an important means to efficiently generate a large number of markers from a desired region of a genome. The present study was designed to initiate microdissection and amplification of DNA from whole chromosomes, arms, or bands of porcine chromosomes. The following pig (SSC) chromosomes/segments were scraped: SSC1p, SSC1q26-q2.13, SSC2q11-q14, SSC4q12-q25, SSC13, SSC13q12-q31, SSC13q32-q43, SSC13q32-q43, SSC15, and SSC16q21-q23. After amplification and PCR-labelling, the DNA from the dissected segments were painted back to normal metaphase chromosomes to test their identity. Microdissection of some of the segments (on SSC4, 13 and 15) coincides with the mapping of economically important traits. As a first step towards generation of markers, microcloning of amplified product from SSC1p and SSC15 was carried out. The libraries were screened with a (GT)15 oligonucleotide probe. Future prospects of such a work in farm animals are discussed.

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Section: Discussionmentioning

confidence: 99%

Microdissection of Pig Chromosomes: Dissection of Whole Chromosomes, Arms and Bands for Construction of Paints and Libraries

et al. 2004

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“…The alignment of QFH bands to that of the standardized karyotype for pig chromosome 6 (data not shown) revealed that the signals were located on band q2z1. were amplified by PCR essentially as described by Grimm et al (1997). The forward and reverse primers flanking the repeat region were 59-AGC TGC TCA GCA CCA AGG TCAC-39 and 59-CTG AGG GTC CAG ACC ACA CGG-39, respectively.…”

Section: Discussionmentioning

confidence: 99%

“…The four alleles (190, 193, 194 and 199 bp) highlighting the position of the microsatellite within the apolipoprotein E (apo-E) gene. The linkage map was developed using the University of Illinois, Chicago, Illinois, USA, reference population and the apo-E gene is located at relative position 91z2 on this map (Clamp et al 1993;Grimm et al 1997;Grimm et al 1998). The cytogenetic map is based on the physical locations of markers in the literature (Yerle et al 1991;Mellink et al 1992;Mellink et al 1993;Paszek et al 1995;Rohrer et al 1996).…”

Section: Discussionmentioning

confidence: 99%

“…These animals represented seven families from this population, selected for their informativeness with framework markers on chromosome 6 (Sw1067, Sw1129, GPI, TGF-B, ENO, PGD, Sw316 and S0031), and additional chromosome 6 markers (DG13, DG30, DG53 and S0444) from the US-MARC and PiGMaP linkage maps (Archibald et al 1995;Paszek et al 1995;Rohrer et al 1996). PCR was performed as described by Grimm et al (1997), but the annealing temperature was 58°C for 1 min. The resulting data were subjected to two-point (LOD > 3z0) linkage analysis with CRIMAP 2z4 (Green et al 1990).…”

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Isolation and genetic characterization of the porcine apolipoprotein E gene

et al. 1998

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The present report describes the isolation and genetic characterization of the porcine apolipoprotein E (apo-E) gene. A single positive recombinant phage clone containing a 10.7-kb insert was isolated from a porcine genomic library, and a 4.2-kb fragment was subcloned and sequenced. The 4.2-kb fragment contained the entire apo-E gene in addition to upstream and downstream sequences (GenBank accession no. 470240). The porcine apo-E gene is made up of four exons and three introns, and encodes a preapo-E protein comprised of a signal peptide of 18 amino acids and a mature protein of 299 amino acids. The porcine apo-E gene contains a (CG)13 microsatellite marker within intron three. This microsatellite is moderately polymorphic, and at least four alleles were evident at this locus among 10 animals from each of the Yorkshire, Hampshire, Landrace and Duroc breeds. Finally, localization of the porcine apo-E gene to chromosome 6 band q2.1 was determined by fluorescent in situ hybridization and confirmed by genetic linkage analysis.

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“…We have previously prepared small insert genomic libraries from swine chromosome 6 DNA isolated by whole chromosome micro‐isolation ( Ambady et al . 1997 ) or bivariate flow sorting ( Grimm et al . 1997 ).…”

Section: Introductionmentioning

confidence: 99%

Microsatellite markers from a microdissected swine chromosome 6 genomic library

Zhao¹,

Ambady²,

León³

et al. 1999

Animal Genetics

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To develop additional microsatellite (MS) markers in the region of the porcine skeletal muscle ryanodine receptor gene (RYR1), a microdissected genomic library was generated from the proximal half of the q arm of swine chromosome 6. Purified DNA was restriction enzyme-digested, ligated to oligonucleotide adaptors and amplified by PCR using primers complementary to the adaptor sequences. The purity of the amplified products and boundaries of the microdissected chromosomal region were verified by fluorescence in situ hybridization. (CA)n-containing sequences were then identified in a small insert genomic library generated from the PCR-amplified microdissected DNA. Oligonucleotide primers were developed for the PCR amplification of 30 of the 46 (CA)n repeat-containing clones, which were subsequently used to amplify DNA isolated from unrelated pigs of different breeds to determine the informativeness of these MS markers. Twenty-two of these MS markers were genotyped on the University of Illinois Yorkshire x Meishan swine reference population. These 22 markers were all assigned within a 50.7-CM region of the swine chromosome 6 linkage map, indicating the specificity of the microdissected library.

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Mapping of microsatellite markers developed from a flow-sorted swine chromosome 6 library

Cited by 9 publications

References 37 publications

Microdissection of Pig Chromosomes: Dissection of Whole Chromosomes, Arms and Bands for Construction of Paints and Libraries

Microdissection of Pig Chromosomes: Dissection of Whole Chromosomes, Arms and Bands for Construction of Paints and Libraries

Isolation and genetic characterization of the porcine apolipoprotein E gene

Microsatellite markers from a microdissected swine chromosome 6 genomic library

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