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Development of a genetic map of the chicken with markers of high utility.

Microsatellites are tandem duplications with a simple motif of one to six bases as the repeat unit. Microsatellites provide an excellent opportunity for developing genetic markers of high utility because the number of repeats is highly polymorphic, and the assay to score microsatellite polymorphisms is quick and reliable because the procedure is based on the polymerase chain reaction (PCR). We have identified 404 microsatellite-containing clones of which 219 were suitable as microsatellite markers. Primers for 151 of these microsatellites were developed and used to detect polymorphisms in DNA samples extracted from the parents of two reference populations and three resource populations. Sixty, 39, 46, 49, and 61% of the microsatellites exhibited length polymorphisms in the East Lansing reference population, the Compton reference population, resource population No. 1 (developed to identify resistance genes to Marek's disease), resource population No. 2 (developed to identify genes involved in abdominal fat), and resource population No. 3 (developed to identify genes involved in production traits), respectively. The 91 microsatellites that were polymorphic in the East Lansing reference population were genotyped and 86 genetic markers were eventually mapped. In addition, 11 new random amplified polymorphic DNA (RAPD) markers and 24 new markers based on the chicken CR1 element were mapped. The addition of these markers increases the total number of markers on the East Lansing genetic map to 273, of which 243 markers are resolved into 32 linkage groups. The map coverage within linkage groups is 1,402 cM with an average spacing of 6.7 cM between loci. The utility of the genetic map is greatly enhanced by adding 86 microsatellite markers. Based on our current map, approximately 2,550 cM of the chicken genome is within 20 cM of at least one microsatellite marker.

Animals↗

High resolution genetic mapping and candidate gene identification at the xa5 locus for bacterial blight resistance in rice ( Oryza sativa L.).

The xa5 resistance gene from rice provides recessive, race-specific resistance to bacterial blight of rice caused by the pathogen Xanthomonas oryzae pv oryzae. A high-resolution genetic map of the chromosomal region surrounding xa5 was developed by placing 44 DNA markers on the distal end of rice chromosome 5. The basis for mapping was a PCR-based screening of 1,016 F(2) individuals derived from a cross between a near-isogenic line (NIL) and its corresponding recurrent parent to identify recombinants in the region. Recombinant F(2) individuals were progeny tested using F(3) families inoculated with the Philippine strain PXO 61 of bacterial blight pathogen. The xa5 gene was mapped to a 0.5-cM interval between the markers RS7 and RM611, which spanned an interval of approximately 70 kb and contained a total of 11 open reading frames. Sequence data for the locus was generated from an Indica (the IR24 isoline, IRBB21) BAC covering part of the region and compared to other overlapping Indica (cv 93-11) and Japonica (cv Nipponbare) sequences. Candidate-gene analysis revealed that a basal transcription factor (TFIIa), an ABC transporter, a tRNA synthase, a MAP kinase and a cysteine protease, as well as four unknown, hypothetical or putative proteins, are encoded at the locus and could be potential candidates for the resistance gene product. The mechanism by which these genes could provide recessive, race-specific resistance will be elucidated by map-based cloning of the xa5 gene.

Chromosomes, Artificial, Bacterial↗

Chiasma-based genetic map of the mouse X chromosome.

The X chromosome pair was identified in diakinesis/metaphase I stage mouse oocytes using a repeat sequence DNA probe and fluorescence in situ hybridisation. Chiasma positions along the X bivalent were measured in 57 oocytes from 4 females. Overall, our observations showed that while there were no obvious "hotspots" for chiasma formation along the X chromosome, there was a tendency to favour the distal end. Minimum inter-chiasma distances were substantial indicating the occurrence of strong genetic interference. Estimates of both genetic distances and recombination fractions for any interval along the chromosome can be calculated from the chiasma data. The average chiasma frequency for the X bivalent was 1.37 giving an estimated total genetic map length of 68.5 cM. In general, the pattern of chiasma distribution along the X chromosome resembled that anticipated from recombination distances in published consensus linkage maps. There were, however, some intriguing differences between the two approaches. The reason for these discrepancies are unknown but may be related to lack of precision in cytogenetic mapping of loci, inter-strain and/or interspecies differences in the genetic controls over the distribution of crossover events. One advantage of the chiasma analysis approach is its suitability for investigating these problems.

Animals↗

Molecular and genetic mapping of the mouse mdx locus.

mdx is an X-linked muscular dystrophy mutant of the mouse and a putative homolog of the human X-linked muscular dystrophy locus--Duchenne muscular dystrophy (DMD). Utilizing a C57BL/10/Mus Spretus interspecific cross in which the mdx mutation was segregating, we have constructed a detailed genetic map around the mdx locus on the mouse X chromosome. We were unable to detect recombinants between mdx and exonic probes derived from the human DMD gene. These genetic data support the contention from biochemical studies (E.P. Hoffman, R. H. Brown, and L. M. Kunkel, 1987, Cell 51: 919-928) that DMD and mdx are homologous genes.

Animals↗

The density distribution of gene loci over the genetic map of Escherichia coli: its structural, functional and evolutionary implications.

A quantitative analysis was carried out on the dispersion of gene loci over the E. coli genetic map. Therefore, the map was divided into regions characterized by an homogeneous gene density. This created a distribution pattern of gene loci that contained a symmetry axis located near to the origin of DNA replication. The pattern could be subdivided into a set of 22 functional domains containing gene loci whose products revealed a biochemical or functional relatedness. A correlation was found between the boundary positions of these domains and the distribution of F plasmid- and DNA insertion sites over the E. coli chromosome. The structural, functional and evolutionary implications of these findings are discussed.

Biological Evolution↗

Genetic mapping of EST-derived microsatellites from the diploid Gossypium arboreum in allotetraploid cotton.

To increase the numbers of microsatellites available for use in constructing a genetic map, and facilitate the use of functional genomics to elucidate fiber development and breeding in cotton, we sampled microsatellite sequences from expressed sequence tags (ESTs) transcribed during fiber elongation in the A-genome species Gossypium arboreum to evaluate their frequency of occurrence, level of polymorphism and distribution in the At and Dt subgenomes of tetraploid cotton. From among ESTs derived from G. arboreum fibers at 7-10 days post anthesis (dpa), 931 ESTs were found to contain simple sequence repeats (SSRs); 544 (58.4%) EST-SSR primer pairs were developed, and 468 (86%) amplified PCR products from allotetraploid cotton (G. hirsutumcv. TM-1 and G. barbadense cv. Hai7124). However, only 99 (18.2%) of these were found to be polymorphic and segregating in our interspecific BC1 mapping population [(TM-1xHai7124)xTM-1]. In these amplified and informative EST-SSRs, hexa- and tri-nucleotide repeat motifs were the most frequent, representing 40.1 and 30%, respectively, of the total. A total of 111 loci detected with these 99 EST-SSRs were integrated into our backbone map including 511 SSR loci. The distribution of the EST-SSRs appeared to be non-random, since 72 loci were anchored to the At and 37 to the Dt subgenome of allotetraploid cotton based on linkage tests. Interestingly, out of the 10 pairs of duplicate loci amplified, seven were mapped to the corresponding homologous linkage groups and/or chromosomes. BLASTX analysis revealed that 69 of the 99 ESTs showed significant similarities to known genes. Some genes important for fiber development, such as sucrose synthase, were mapped to corresponding chromosomes. These EST-SSRs provide structural and functional genomic information that will be useful for understanding cotton fiber development.

Base Sequence↗

CoIV plasmids pCoIV-B188 and pCoIV-K30: genetic maps according to restriction enzyme sites and landmark phenotypic characteristics.

The CoIV plasmids are large virulence plasmids of the incompatibility group IncFI. We have obtained the genetic maps of two of the most studied CoIV plasmids, pCoIV-B188 and pCoIV-K30, according to restriction enzyme sites and landmark phenotypic characteristics such as colicin V, the aerobactin iron uptake system, the transfer region, replication regions, and repeated sequences. Although the two plasmids differ in size (pCoIV-B188 is 80 kb and pCoIV-K30 is 144 kb), the maps reveal many regions which are apparently identical or very similar.

Escherichia coli↗

[Computer program of genetic map construction and database of gene localization].

Computer data base containing information about gene localization in the chromosomes of various organisms is composed. Computer program of genetic map constructing is created. The program represents graphically the information conserving in the data base. The data base and the program are intended for realization on the IBM-compatible computers.

Chromosome Mapping↗

The use of pedigree, sib-pair and association studies of common diseases for genetic mapping and epidemiology.

Efforts to identify gene variants associated with susceptibility to common diseases use three approaches: pedigree and affected sib-pair linkage studies and association studies of population samples. The different aims of these study designs reflect their derivation from biological versus epidemiological traditions. Similar principles regarding determination of the evidence levels required to consider the results statistically significant apply to both linkage and association studies, however. Such determination requires explicit attention to the prior probability of particular findings, as well as appropriate correction for multiple comparisons. For most common diseases, increasing the sample size in a study is a crucial step in achieving statistically significant genetic mapping results. Recent studies suggest that the technology and statistical methodology will soon be available to make well-powered studies feasible using any of these approaches.

Chromosome Mapping↗

[Genetic map and structure in "Escherichia coli" K12 of a resistance plasmid isolated from "Salmonella ordonez" (author's transl)].

A resistance plasmid called R IP173 has been transferred into E. coli K12 from a multiresistant strain of S. ordonez isolated during an epidemic in Dakar. This plasmid mediates for colicine Ib production and resistance to ampicillin, streptomycin, spectinomycin, kanamycin, chloramphenicol, tetracycline and sulfonamides. It is transducible "en bloc" by the phage P1-kc between strains of E. coli K12. Compatibility studies have shown that R IP173 belongs to the fi- class, I1 group. It is transferred "en bloc" in conjugation experiments between E. coli K12 strains. But during transfers from S. ordonez into E. coli, incomplete variants are obtained, lacking different markers. A deletion map was obtained after analysis of 19 different variants, and it is suggested that the loss of markers results from the loss of genetic material during transfer. In this particular case, the deletions observed in transduction or conjugation experiments lead to identical genetic maps.

Ampicillin↗

Genetic mapping of antibiotic resistance in markers Bacillus subtilis.

Several antibiotic resistance markers in Bacillus subtilis have been mapped by three-factor transduction crosses using bacteriophage PBS1. These markers, which are thought to be located in genes coding for ribosomal proteins, are within a segment comprising about 5 per cent of the B. subtilis genetic map, close to the major group of rRNA cistrons. The order of genetic makers in this region was found to be cysA14 mic-1 dagger str-1 (ery-1, ole-2) spc-2 bry-2 lin-2. The positions of neo-2, nea-1, and kan-2 are uncertain, although they are located in this region to the right of cysA14.

Anti-Bacterial Agents↗

Ornithine decarboxylase in Saccharomyces cerevisiae: chromosomal assignment and genetic mapping of the SPE1 gene.

The gene for ornithine decarboxylase in Saccharomyces cerevisiae, SPE1, has been assigned to chromosome XI by the technique of transverse alternating pulsed field electrophoresis and DNA-DNA hybridization. Genetic mapping by tetrad analysis shows that the SPE1 gene is located on the left arm of chromosome XI, 6 cM from the LAP1 gene and 43 cM from the TRP3 gene. The spe10 mutation previously isolated in this laboratory is mapped to the N-terminal region of the SPE1 gene, and therefore should be designated as a spe1 allele.

Chromosome Mapping↗

Genetic mapping of the Salmonella typhimurium pepB locus.

Transposon technology has been used to map the pepB locus of Salmonella typhimurium. This locus is cotransducible by phage P22 with glyA and strB at min 56 on the Salmonella genetic map. The gene order is strB pepB glyA.

Aminopeptidases↗

Attempts to extend the genetic map of poliovirus temperature-sensitive mutants.

Eighteen new ts mutants of poliovirus have been isolated after a variety of mutagenic treatments, and their loci identified in relation to the previous genetic map. The map was only extended by 25%, and the physiological characters of the new isolates corresponded in all aspects tested with those of the previous isolates. Apparently single mutants at the extreme left of the map were defective in synthesis of both double- and single-stranded RNA, functions that do not co-vary in other mutants. Two procedures respectively predicted to induce mutations preferentially in the 5' and 3' regions of the genome gave isolates which all indicated that the structural protein region was nearest the 5' end. The loci fro resistance to dextran sulphate and to ethyl-2-methylthio-4-methyl-5-pyrimidine carboxylate both lie in the structural protein region.

Chromosome Mapping↗

Detailed physical and genetic mapping in the region of plasminogen, D17Rp17e, and quaking.

We present here a detailed physical map encompassing over 600 kb of mouse Chromosome (Chr) 17 in the region of plasminogen, D17Rp17e, and quaking. This region is cloned in yeast artificial chromosomes (YACs). We have identified several CpG islands within this region from pulsed field gel mapping of mouse genomic DNA and YAC DNA. Five new DNA probes have been generated. One, D17Leh514, is a minimum of about 90 kb distal to plasminogen. Four, D17Leh513, D17Leh512, D17Leh511, and D17Leh510, are proximal to D17Rp17e, the closest previously described genetic marker to quaking viable and quaking lethal-1 mutations. We have genetically mapped D17Leh511 to within 0.15 cM of these mutations. The genetic distance to D17Rp17e from D17Leh511 is also 0.15 cM; the physical distance of less than 360 kb (minimum 200 kb) is consistent with an approximation of 2 Mbp per cM.

Animals↗

Genetic mapping of the alpha-galactosidase MEL gene family on right and left telomeres of Saccharomyces cerevisiae.

The alpha-galactosidase MEL2-MEL10 genes have been genetically mapped to right and left telomere regions of the following chromosomes of Saccharomyces cerevisiae: MEL2 at VII L, MEL3 at XVI L, MEL4 at XI L, MEL5 at IV L, MEL6 at XIII R, MEL7 at VI R, MEL8 at XV R, MEL9 at X R and MEL10 at XII R. A set of tester strains with URA3 inserted into individual telomeres and no MEL genes was used for mapping.

Chromosome Mapping↗

Molecular cloning and genetic mapping of the t complex responder candidate gene family.

Male transmission ratio distortion (TRD) is a property of mouse t haplotypes requiring the t complex responder locus (Tcr). Tcr maps to the central region of t haplotypes, and is embedded within a series of large duplicated tracts of DNA known as "T66 elements." In previous work, a family of genes (the "T66" genes) was identified within this region that encodes male germ cell-specific transcripts. Genetic and molecular data indicate that one of these genes represents Tcr. Here, we describe the molecular cloning of the four members of the T66 gene family, the genetic mapping of these genes to three adjacent t haplotype loci, and comparative restriction enzyme analysis of the genes. The results indicate that these genes are highly similar to one another, and were created by recent, complex duplication events. This suggests that a minor alteration(s) could have been responsible for conferring "mutant" responder activity upon Tcr, while the other homologs retained "wild-type" biochemical function. In addition, we have identified and mapped three T66 genes in wild-type t complexes. They reside in two separate loci at the opposite ends of the proximal t complex inversion, and are separated by at least 3 cM.

Animals↗

Physical and genetic maps of the Leptospira interrogans serovar icterohaemorrhagiae strain Ictero no.1 chromosome and sequencing of a 19-kb region of the genome containing the 5S rRNA gene.

We report the construction of physical and genetic maps of the chromosome of Leptospira interrogans serovar icterohaemorrhagiae strain Ictero No.1 using pulsed-field gel electrophoresis of DNA fragments generated by digestion with enzymes SrfI, AscI, FseI, and NotI and using reciprocal hybridization. We also sequenced the 19-kilobase (kb) DNA segment including the one gene for 5S rRNA (rrf) of pathogenic Leptospira. The size of the chromosome of the strain Ictero No.1 was estimated to be 4673kb and was found to be similar to those of the chromosomes of the leptospira strains Verdun (serovar icterohaemorrhagiae) and RZ11 (serovar pomona). The strains Verdun and RZ11 carry a small 350-kb replicon (minichromosome), and the strain Ictero No.1 also contained the same kind of molecule together with the chromosome. The physical maps of the strains Ictero No.1 and Verdun were almost identical, as were the locations of the selected genes, except for the location of one of the 16S rRNA genes. Overall, the genetic organization appeared to be conserved within the serovar icterohaemorrhagiae strains. In the sequenced region, we identified 10 putative ORFs and one rrf sequence, and the transcription orientations were all the same. A homology search for the products deduced from the sequenced data revealed that the orf H exhibited high similarity to malic acid enzyme of Haemophilus influenzae and fumarate hydratase of Escherichia coli (orf J). The rest of the putative products encoded by ORFs in the sequenced region showed little similarity with the proteins contained in the databases and were considered to be unknown proteins.

Bacterial Proteins↗