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Biochemical Genetics (v.40, #9-10)

Haplotype Diversity of the Mitochondrial DNA D-Loop Region in Calomys musculinus (Rodentia, Muridae) Detected by PCR-RFLP by Raúl E. González Ittig; Cristina N. Gardenal (pp. 293-302).

In order to contribute to knowledge of colonization patterns in the rodent Calomys musculinus, a natural reservoir of the virus producing Argentine hemorrhagic fever (AHF), we studied the haplotype diversity of the mitochondrial DNA D-loop region in five natural populations from central Argentina. Digestion with eight restriction enzymes (RsaI, MseI, Tsp509I, AluI, AciI, HaeIII, NlaIII, and AseI) revealed polymorphism in the 1300 bp fragment amplified by PCR. Twenty different composite haplotypes were detected. Hierarchical analyses indicated that almost all variation (94%) is contained within local populations. Haplotypes 1 and 2, shared by all populations, were the most frequent. Nonsignificant genetic differentiation was found among populations of the endemic and nonendemic areas of AHF. All locations sampled presented exclusive haplotypes in spite of their geographic proximity, which would support previous observations indicating restricted gene flow among C. musculinus populations.

Keywords: Calomys musculinus ; RFLP-PCR; haplotype diversity; D-loop region; Argentine hemorrhagic fever

Cloning and Identification of a Novel cDNA¹ Which May Be Associated with FKBP25 by Xianfang Meng; Jinzhong Chen; Quansheng Yang; Shu Wang; Yaoqiong Chao; Kang Ying; Yi Xie; Yumin Mao (pp. 303-310).

A 1209 bp novel cDNA with a putative open reading frame of 627 bp that has an overlapped fragment of 527 bp with FKBP25 was isolated from a human fetal brain library. Northern Blot analysis detected a signal about 1.5 kb that suggests it is a full-length cDNA. Reverse transcription-PCR demonstrates that the transcripts are high in adult brain, testis, ovary, spleen, and fetal brain. Comparing the cDNA with the human genome database identifies a contig that reveals that the gene contains six exons and is located on human chromosome 20p11. The gene encodes a putative protein with an unknown domain conserved in many species and is similar to bacterial histidyl-tRNA synthetase, and so it is named human histidyl-tRNA-synthetase-related gene.

Keywords: human histidyl-tRNA-synthetase-related gene; FKBP25; Northern Blot; reverse transcription-PCR

Genetic Analysis of Jumbled Spine and Ribs (Jsr) Mutation Affecting the Vertebral Development in Mice by Shinya Okano; Atsushi Asano; Yasuhiro Kon; Hiroyuki Miyoshi; Tomomasa Watanabe (pp. 311-322).

The jumbled spine and ribs (Jsr) mouse was derived from a spontaneous mutation. As the phenotype, a shortened trunk and kinky tail are characteristic Jsr traits. In this study, on high resolution mapping it was found that Lunatic fringe (Lfng) mapped at the same position as Jsr. Lfng was identified as the candidate gene for Jsr, but sequence analysis of this gene revealed no substitution in the coding region of cDNA. Therefore, we adopted the strategy of positional cloning for Jsr using a mouse bacterial artificial chromosome (BAC) library. A BAC contig was constructed from three BAC clones showing positive signals of Lfng and 11MMHAP75FRD8.seq near the Jsr locus on chromosome 5. Based on the genetic mapping of both T7 and sp6 ends of a clone of BAC382-O-7 (BAC382), the Jsr gene was considered to exist in BAC382 and to be positioned near the sp6 side.

Keywords: Jsr ; vertebral development; Lfng ; BAC

Assessment of Genetic Structure of Greek Brown Hare (Lepus europaeus) Populations Based on Variation in Random Amplified Polymorphic DNA (RAPD) by Zissis Mamuris; Athanasios I. Sfougaris; Costas Stamatis; Franz Suchentrunk (pp. 323-338).

The RAPD method was used to assess the genetic differentiation of brown hare (Lepus europaeus) populations from Central Greece. Greek wild populations were compared with samples from Austria, Poland, Germany, France, and Bulgaria, as well as with reared/released hares to investigate the impact of the releases on the native populations' genetic structure. The absence of diagnostic bands distinguishing between L. europaeus populations confirmed the high level of gene flow between brown hare populations over long geographic distances reported by other authors. Phylogenetic trees, derived from genetic distances estimated by RAPD band frequencies, suggested one major partitioning event of nuclear DNA lineages found in the samples. The reared individuals clustered with the Austrian, Polish, German, and French populations, whereas the Greek populations clustered apart with the Bulgarian population. Within Greece the distribution of the six wild populations did not follow any geographical trend, since their genetic divergence did not seem to correlate to geographic distances. However, RAPD profiles of some reared and wild specimens were different from the common RAPD pattern observed in the vast majority of sampled hares, probably reflecting an admixture of genetically differentiated individuals. The RAPD analysis indicates that releases might have begun to affect Greek population structure and reinforces the view that appropriate management is needed, adjusted to the local populations' biology and ecology.

Keywords: Lepus europaeus ; genetic diversity; genetic conservation; RAPDs

Genetic and Phenotypic Polymorphisms of the A Subunit of Coagulation Factor XIII in Japanese Population by Masato Taniguchi; Tokiyoshi Ayabe; Toshifumi Ashida; Yoshinori Fujimoto; Yutaka Kohgo (pp. 339-349).

To clarify whether genetic polymorphisms in exon 14 of Coagulation factor XIII A-subunit gene (FXIIIA) affect phenotype expressions, we studied genetic polymorphisms in exon 14 of FXIIIA in a Japanese population and the relationship between the genetic polymorphisms and phenotype expression. Genetic polymorphisms in exon 14 of FXIIIA of 144 unrelated Japanese were analyzed by single-strand conformation polymorphism. Plasma FXIIIA antigen concentrations, FXIII activities, and phenotype were also determined by two-dimensional electrophoresis. The frequencies of the three genotypes, the homozygote (AD), the homozygote (BC) and the heterozygote (AD/BC), were 77.1, 0.7, and 22.2%, respectively. The gene frequencies of AD and BC were 0.88 and 0.12. We detected AD (GTTand BC (ATTat codon 650 and 651 of exon 14. There were no significant differences of FXIIIA antigen concentrations and FXIII activities between these genotypes. We detected three pl differences among them as being pls of 5.3, 5.6, 5.8 in the homozygote (AD) and the heterozygote (AD/BC), and a pl of 5.8 in the homozygote (BC). These polymorphisms affected isoelectric mobility, but did not affect protein levels, enzyme activities, or the molecular weight of FXIII.

Keywords: factor XIII activity; factor XIIIA antigen concentration; factor XIIIA polymorphism; two-dimensional electrophoresis

RAPD Markers to Evaluate Callus Tissue of Cereus peruvianus Mill. (Cactaceae) Maintained in Different Growth Regulator Combinations by Claudete A. Mangolin; Laura M. M. Ottoboni; Maria de Fátima P. S. Machado (pp. 351-358).

RAPD markers were used to detect DNA polymorphisms in callus tissues maintained at different auxin and cytokinin combinations. There is a higher level of genetic variablity in callus tissue maintained with the highest kinetin versus2, 4-D concentration. Callus tissues subcultured in a 4.0 mg/L 2,4-D and 4.0 mg/L kinetin combination showed high similarity and can be recommended as more suitable sources for industrial procedures of extraction of natural products such as secondary metabolites since extraction protocols can be easily standardized using genetically uniform materials. The higher genetic diversity in callus tissues of C. peruvianus cultured at 4.0 mg/L 2,4-D and 8.0 mg/L kinetin indicates this tissue as a matrix for in vitro selection of cell lines for higher natural products production. RAPD markers are, therefore, effective tools useful for detecting DNA polymorphism in callus tissue as well as in the DNA identification of callus tissues maintained in different auxin and cytokinin combinations.

Keywords: RAPD markers; callus tissue; cactus; tissue culture; growth regulators

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Biochemical Genetics (v.40, #9-10)

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Biochemical Genetics (v.40, #9-10)