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Biochemical Genetics (v.43, #1-2)
Detection of p16 Hypermethylation in Gastric Carcinomas Using a Seminested Methylation-Specific PCR by Peng Hou; Mei-Ju Ji; Jia-Yao Shen; Nong-Yue He; Zu-Hong Lu (pp. 1-9).
Aberrant DNA methylation of a CpG site is among the earliest and most frequent alterations in various tumors including gastric carcinoma. The aim of this study is to detect tumor-associated aberrant hypermethylation of the p16 gene from 60 gastric tumor and corresponding normal tissues using a seminested methylation-specific PCR (MSP). The results indicated that hypermethylation of the p16 gene could be detected in 80% (48/60) of the gastric tumor samples from the first PCR. However, the frequency increased significantly to 86.7% (52/60) of the gastric tumor samples after the second PCR. These results show that this technique increases the sensitivity of detecting p16 hypermethylation from tumor samples. Furthermore, the aberrant methylation of p16 was observed in all of the stages, confirming that this epigenetic alteration is an early event during gastric carcinogenesis. Clinicopathologic parameters such as age, sex, and histological differentiation of GC were not significantly associated with the methylation status.
Keywords: p16 gene; hypermethylation; gastric carcinomas; seminested MSP
Allozymic Polymorphism Among 14 Populations of the House Mouse, Mus musculus domesticus, From Greece by Georgios Tryfonopoulos; Basil Chondropoulos; Stella Fraguedakis-Tsolis (pp. 11-24).
Nineteen loci from 239 individuals of the house mouse Mus musculus domesticus (Rodentia, Muridae) were analyzed by means of thin layer electrophoresis. The mice were collected from 14 localities of Greece mainly confined to the area of NW Peloponnese, where a Robertsonian (Rb) system is observed. The individuals were chromosomally characterized by nine diploid numbers, the 2n = 24, 26, 27, 28, 29, 30, 31, 32, and 40. The statistic elaboration revealed that all 14 populations studied were not characterized by cohesive demic structure and high inbreed levels while the gene flow among them has resulted in low levels of genetic differentiation. The resulting values for Nei’s genetic distance corresponded to distances known for the level of geographical populations of, M. musculus. Wagner’s cladogram for the phylogenetic relations between the populations studied implied that it is the diploid number, rather than the geographical factor, that characterizes or dominates each population, which mainly influences the phylogenetic relationships.
Keywords: Mus musculus domesticusRb populations; allozymes; genetic polymorphism; Greece
Molecular Genetic Analysis of QTLs for Ferulic Acid Content in Dried Straw of Rice (Oryza sativa L.) by Yanjun Dong; E. Tsuzuki; H. Kamiunten; Dongzhi Lin; H. Terao; M. Matsuo; Shihua Cheng (pp. 25-34).
Phenolic acids are secondary metabolic organic compounds produced by plants and often are mentioned as allelochemicals. This study was conducted to determine the genetic basis controlling the ferulic acid content of rice straw in a recombinant inbred (RI) population derived from a cross between a japonica variety, Asominori, with a higher content of ferulic acid, and an indica variety, IR24, with a lower content, using 289 RFLP markers. Continuous distributions and transgressive segregations of ferulic acid content were observed in the RI population, which showed that ferulic acid content in rice straw was quantitatively inherited. Single marker analysis and composite interval mapping identified three quantitative trait loci (QTLs) for ferulic acid content with LOD values of 2.03 (chromosome 3), 3.16 (chromosome 6), and 3.06 (chromosome 7); all three had increased additive effects (13.5, 18.3, and 18.1 μg g −1) from the Asominori parent and accounted for 5.5, 16.9, and 12.8% of total phenotypic variation, respectively. This is the first report on the identification of QTLs associated with ferulic acid and their chromosomal localization on the molecular map of rice. The tightly linked molecular markers that flank the QTLs might be useful in breeding and selection of varieties with higher phenolic acid content.
Keywords: ferulic acid; QTLs; molecular markers; rice straw
Molecular Analysis of the Relatedness of Five Domesticated Turkey Strains by Edward J. Smith; Tuoyu Geng; Elizabeth Long; F. William Pierson; D. Phillip Sponenberg; Cal Larson; Robert Gogal (pp. 35-47).
Our knowledge of the genetic relatedness among the eight existing domesticated turkey strains is limited. To begin to address this paucity, genetic relatedness among five turkey strains (Blue Slate, Bourbon Red, Narragansett, Royal Palm, and Spanish Black) was investigated using three molecular marker systems: randomly amplified polymorphic DNA (RAPD), microsatellite, and SNPs derived from a sequence tagged site and a cloned RAPD fragment. The RAPD analyses were based on five primers that revealed a total of 14 informative DNA fragments in all five populations. The microsatellite analyses involved two informative alleles from three primer-pairs. A total of nine SNPs were detected, one of which appeared to be strain specific. This SNP formed the basis of a PCR-RFLP genotyping procedure developed to distinguish one of the strains from the other four. Evidence from these analyses including the SNP-based RFLP-PCR suggests that Royal Palm is distinct from the other four strains, though more closely related to Narragansett. These data provide, for the first time, molecular evidence of the potential relationships among noncommercial domesticated turkey strains.
Keywords: turkeys; genetic relatedness; molecular analysis
Isoenzyme Variation in Melipona rufiventris (Hymenoptera: Apidae, Meliponina) in Minas Gerais State, Brazil by Ronaldo Guimarāes Costa; Mara Garcia Tavares; Luiz Antonio dos Santos Dias; Lucio Antonio de Oliveira Campos (pp. 49-58).
The stingless bee Melipona rufiventris is an important pollinator in several Brazilian ecosystems. Originally widely distributed in Minas Gerais (MG) state, this species is becoming very rare. Therefore this species was included in the endangered species list of MG. We used isoenzyme data for a better understanding of the genetic structure of several M. rufiventris colonies. Samples of 35 colonies were collected from 12 localities and evaluated by nine enzymatic systems, which yielded 17 loci. M. rufiventris genetic variation was found to be low, typical of an endangered species. The proportion of polymorphic loci was 5.88% in both ecosystems. Only Est-4 was polymorphic in colonies from the Forest and Mdh-1 in colonies from the Cerrado. The expected heterozygosity ranged from 0.0068 in the Cerrado to 0.0078 in the Forest. Despite this, enzyme electrophoretic analyses provided a good idea of the diversity between samples from Cerrado and Forest which reinforce the existence of two different “forms” of M. rufiventris in MG, one present in the Cerrado and the other in Forest. This information is of great importance for the conservation of, M. rufiventris in MG.
Keywords: HymenopteraMeliponagenetic variability; isozymes
Cloning and Characterization of a β3 Tubulin cDNA From the Small Brown Planthopper, Laodelphax striatellus
by Jian-Guo Wen; Jian Yan; Jia Xu; Da-Leng Shen (pp. 59-64).
Defective Glycosaminoglycan Substitution of Decorin in a Patient With Progeroid Syndrome Is a Direct Consequence of Two Point Mutations in the Galactosyltransferase I (ß4galT-7) Gene by Martin Götte; Hans Kresse (pp. 65-77).
The small dermatan sulfate proteoglycan decorin is involved in the regulation of collagen fibrillogenesis, cell adhesion and migration, and growth factor signaling. In a progeroid patient carrying two point mutations in ß4galactosyltransferase I (ß4galT-7) only 50% of the decorin core protein molecules are substituted with glycosaminoglycan chains. We expressed decorin, as well as wild-type and mutant alleles of ß4galT-7 in galactosyltransferase-deficient CHO618 cells. Decorin was less efficiently substituted with glycosaminoglycan chains upon expression of ß4galT-7186D compared to ß4galT-7-expressing cells. Decorin from ß4galT-7-expressing cells displayed increased molecular heterogeneity. Decorin glycosaminoglycan chains were completely susceptible to chondroitinase ABC treatment. Cells expressing ß4galT-7206P did not synthesize the proteoglycan form of decorin. Thus, the ß4galT-7 mutations directly affect the molecular phenotype of decorin observed in a patient with the progeroid form of Ehlers-Danlos syndrome, which may be a major mechanistic cause for the skin and wound healing defects observed in this patient.
Keywords: galactosyltransferase deficiency; decorin; glycosaminoglycan; proteoglycan; dermatan sulfate
Assignment of the Gene for Porcine Insulin-Like Growth Factor Binding Protein 1 to Chromosome 18 and Detection of Polymorphisms in Intron 2 by PCR–RFLP by Yuko Inage-Miyake; Shin-ichi Shimanuki; Tomohito Itoh; Yasuto Murakami; Masahiro Kimura; Hideaki Suzuki; Masashi Miyake; Daisuke Toki; Hirohide Uenishi; Takashi Awata; Noriyuki Hamasima (pp. 79-85).
We have obtained a partial cDNA and three BAC clones for the porcine insulin-like growth factor binding protein 1 gene (IGFBP-1). Results of fluorescence in situ and radiation hybrid (RH) mapping assigned this gene to porcine chromosome (SSC) 18q24-qter. We found two types of polymerase chain reaction–restriction-fragment-length polymorphisms (PCR–RFLP) in intron 2 by using FokI and AluI.
Keywords: IGF-binding proteins I; IGFBPs; FISH; RH mapping; polymorphisms